Phospho-specific antibodies’ typical purpose is to enable researchers to detect changes in proteins. They will exclusively bind to the amino acid sequence on a protein that has been phosphorylated (which is both a physical & chemical change) and do not bind to the same amino acid sequence on said protein if it lacks said phosphorylation. This aids in being able to clearly see and understand the data produced from this particular protein modification.
IHC (Immunohistochemistry) (At 1/100 staining Human colorectal cancer and adjacent normal tissues by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human colorectal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human kidney cancer and adjacent normal tissues by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human kidney cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human ovarian cancer and adjacent normal tissues by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse muscle tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistchemistry) (At 1/100 staining Rat liver tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat spleen tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistchemistry) (At 1/100 staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat stomach tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of POLR2A (Phospho-Ser1616) using Forskolin treated 293 whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
WB (Western Blot) (Western blot analysis of extracts from 293, rat brain and HepG2, using POLR2A (Phospho-Ser1616) Antibody.)
Human, Mouse, Rat Predicted Reactivity: Pig (100%), Zebrafish (100%), Bovine (100%), Horse (100%), Sheep (100%), Dog (100%), Xenopus (100%)
Applications
Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Application Data (At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human ovarian cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse colorectal tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of TSC2 (Phospho-Ser1254) using COLO205 whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Application Data (At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse liver tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Caspase 3 (Phospho-Ser26) using Rat kidney tissue lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Human, Mouse, Rat Predicted Reactivity: Rabbit (88%)
Applications
Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
WB (Western Blot) (Western blot analysis of ENO3 over-expressed 293 cell line, cotransfected with ENO3 Validated Chimera RNAi (Lane 2) or non-transfected control (Lane 1). Blot probed with ENO3 monoclonal antibody. GAPDH (36.1kD) used as specificity and loading control.)
Application Data (Detection limit for recombinant GST tagged ENO3 is ~0.03ng/ml as a capture antibody.)
IF (Immunofluorescence) (Immunofluorescence of monoclonal antibody to ENO3 on HeLa cell. [antibody concentration 10ug/ml].)
IHC (Immunohistochemistry) (Immunoperoxidase of monoclonal antibody to ENO3 on formalin-fixed paraffin-embedded human colon. [antibody concentration 3ug/ml].)
IHC (Immunohistochemistry) (Immunoperoxidase of monoclonal antibody to ENO3 on formalin-fixed paraffin-embedded human stomach. [antibody concentration 1.5ug/ml].)
WB (Western Blot) (Western Blot analysis of ENO3 expression in transfected 293T cell line by ENO3 monoclonal antibody. Lane 1: ENO3 transfected lysate (46.9kD). Lane 2: Non-transfected lysate.)
WB (Western Blot) (Western Blot detection against Immunogen (31.24kD).)
IHC (Immunohistchemistry) (At 1/100 staining Human colorectal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human ovarian cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of CDH1 (Phospho-Ser844) using PI3-kina H2O2 treated COS7 whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
IHC (Immunohistochemistry) (At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistchemistry) (At 1/100 staining Rat spleen tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis IRF-7 (Phospho-Ser477) using Serum treated LOVO whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
WB (Western Blot) (Detection of human phospho KAP-1 (S824) by western blot. Samples: Whole cell lysate (10 ug) from HEK293T cells treated with 100 uM etoposide (+) or mock treated (-). Antibody: Affinity purified rabbit anti-phospho KAP-1 (S824) antibody (AAA23773 lot 6) used for WB at 0.04 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds. Lower Panel: Rabbit anti-KAP1 antibody .)
WB (Western Blot) (Detection of mouse phospho KAP-1 (S824) by western blot. Samples: Whole cell lysate (10 ug) from NIH 3T3 cells treated with 100 uM etoposide (+) or mock treated (-). Antibody: Affinity purified rabbit anti-phospho KAP-1 (S824) antibody (AAA23773 lot 6) used for WB at 0.04 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds. Lower Panel: Rabbit anti-KAP-1 antibody .)
IP (Immunoprecipitation) (Detection of human phospho KAP-1 (S824) by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 10% of IP loaded) from HEK293T cells treated with 100 uM etoposide (+) or mock treated (-). Antibodies: Affinity purified rabbit anti-phospho KAP-1 (S824) antibody (AAA23773 lot 6) and rabbit anti-KAP-1 recombinant monoclonal antibody used for IP at 6 ug per reaction. For blotting immunoprecipitated Phospho KAP-1 (S824), AAA23773 was used at 0.04 ug/ml. Detection: Chemiluminescence with an exposure time of 1 second. Lower Panel: Rabbit anti-KAP1 antibody .)
IHC (Immunohistochemistry) (Detection of human Phospho KAP-1 (S824) by immunohistochemistry. Samples: FFPE serial sections of prostate carcinoma with MOCK CIP treatment (left) and prostate carcinoma with CIP treatment (right). Antibody: Affinity purified rabbit anti-Phospho KAP-1 (S824) (Cat. No. AAA23773 lot 6) used at a dilution of 1:200 (1 ug/ml). Detection: DAB)
ICC (Immunocytochemistry) (Detection of human Phospho KAP-1 (S824) by immunocytochemistry. Samples: NBF-fixed asynchronous HeLa cells grown in chambered microscope slides and treated with etoposide (left) or untreated (right). Antibody: Affinity purified rabbit anti-Phospho KAP-1 (S824) (Cat. No. AAA23773) used at a dilution of 1:200 (1ug/ml). Detection: Red fluorescent anti-rabbit IgG-DyLight 594 conjugated used at a dilution of 1:100.)
ICC (Immunocytochemistry) (Detection of human Phospho KAP-1 (S824) by immunocytochemistry. Samples: FFPE serial sections of asynchronous etoposide treated HeLa cells with MOCK CIP treatment (left) and etoposide treated HeLa cells with CIP treatment (right). Antibody: Affinity purified rabbit anti-Phospho KAP-1 (S824) (Cat. No. AAA23773 lot 6) used at a dilution of 1:200 (1 ug/ml). Detection: DAB)
IHC (Immunohistochemistry) (At 1/100 staining Human esophageal cancer and adjacent normal tissues by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistchemistry) (At 1/100 staining Human esophageal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human esophageal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse kidney tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of eIF2B (Phospho-Ser2) using Mouse brain tissue lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
IHC (Immunohistchemistry) (At 1/100 staining Mouse liver tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat heart tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis NDEL1 (Phospho-Thr219) using UV treated Jurkat whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Application Data (At 25 degree C. Samples were then incubated with primary Ab(At 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)
IHC (Immunohistchemistry) (At 1/100 staining Human colorectal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human pancreatic cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of PDHA1/2 (Phospho-Ser293/291) using HeLa whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
IF (Immunofluorescence) (Immunofluorescence analysis of U-2 OS cells using Phospho-Histone H3-S10/T11 Rabbit pAb at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IF (Immunofluorescence) (Immunofluorescence analysis of NIH-3T3 cells using Phospho-Histone H3-S10/T11 Rabbit pAb at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IF (Immunofluorescence) (Immunofluorescence analysis of C6 cells using Phospho-Histone H3-S10/T11 Rabbit pAb at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded mouse kidney using Phospho-Histone H3-S10/T11 antibody at dilution of 1:100 (40x lens).)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded rat liver using Phospho-Histone H3-S10/T11 antibody at dilution of 1:100 (40x lens).)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded human lung cancer using Phospho-Histone H3-S10/T11 antibody at dilution of 1:100 (40x lens).)
WB (Western Blot) (Western blot analysis of extracts of NIH/3T3 cells, using phospho-STK4-T387 pAb at 1:1000 dilution or Histone H3 antibody (A15741).HeLa, NIH/3T3 and C6 cellls were treated by nocodazole (50 ng/mL) at 37℃ for 20 hours.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit (RM00020).Exposure time: 1s.)
WB (Western Blot) (Western blot analysis of extracts of various cell lines, using Phospho-Histone H3-S10/T11 antibody at 1:1000 dilution. Both NIH/3T3 cells and Hela cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour and treated by nocodazole (50 ng/mL) at 37℃ for 20 hours.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% BSA.Detection: ECL Basic Kit (RM00020).Exposure time: 1s.)
IF (Immunofluorescence) (Immunofluorescence analysis of U-2 OS cells using Phospho-Histone H3-T6 Rabbit pAb (AAA28403) at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IF (Immunofluorescence) (Immunofluorescence analysis of NIH-3T3 cells using Phospho-Histone H3-T6 Rabbit pAb (AAA28403) at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IF (Immunofluorescence) (Immunofluorescence analysis of C6 cells using Phospho-Histone H3-T6 Rabbit pAb (AAA28403) at dilution of 100 (40x lens). Blue: DAPI for nuclear staining.)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded mouse pancreas using Phospho-Histone H3-T6 antibody (AAA28403) at dilution of 1:100 (40x lens).)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded mouse testis using Phospho-Histone H3-T6 antibody (AAA28403) at dilution of 1:100 (40x lens).)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded rat brain using Phospho-Histone H3-T6 antibody (AAA28403) at dilution of 1:100 (40x lens).)
IHC (Immunohistochemistry) (Immunohistochemistry of paraffin-embedded human colon using Phospho-Histone H3-T6 antibody (AAA28403) at dilution of 1:100 (40x lens).)
WB (Western Blot) (Western blot analysis of extracts of various cell lines, using Phospho-Histone H3-T6 antibody (AAA28403) at 1:1000 dilution.Hela cells and NIH/3T3 cells were treated by CIP(20uL/400ul) at 37 degree C for 1 hour.HeLa cells were treated by nocodazole (50 ng/ml) at 37 degree C for 20 hours.Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.Lysates/proteins: 25ug per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Enhanced Kit (RM00021).Exposure time: 1s.)
IHC (Immunohistchemistry) (Immunohistochemical analysis of (AAA28653) on paraffin-embedded Human breastcarcinoma tissue. Tissue was fixed withformaldehyde at room temperature. Heatinduced epitope retrieval was performed byEDTA buffer (pH9. 0). Samples wereincubated with primary antibody(1:100) for 1hour at room temperature. Undiluted CRFAnti-Polyvalent HRP Polymer antibody wasused as the secondary antibody.)
WB (Western Blot) (Western blot analysis of lysates from A2780 cell line, untreated or treated with Alkaline phosphatase, 1h, using 459085101 (AAA28653) or Beta-actin (lower).)
WB (Western Blot) (Western blot analysis of lysates from A2780 cell line, untreated or treated with Alkaline phosphatase, 1h, using 459088101 (AAA28653) (upper) or Beta-actin (lower).)
WB (Western Blot) (Western blot analysis of lysates from 293 cell line, untreated or treated with Alkaline phosphatase, 1h, using 459088102(AAA28653) (upper) or Beta-actin (lower).)
IHC (Immunohistochemistry) (Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.)
WB (Western Blot) (The anti-phospho CDC25A S124 Pab is used in Western blot to detect CDC25A in mouse liver tissue lysate)
IHC (Immunohistchemistry) (At 1/100 staining Rat heart tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat skin tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistchemistry) (At 1/100 staining Human esophageal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Rat spleen tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human colorectal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
IHC (Immunohistochemistry) (Staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of VR1(Phospho-Ser502) using NIH-3T3 whole cell lysates.-/+ means absence or presence of N peptide(non-phospho peptide) and P peptide(phospho peptide).)
Human, Mouse, Rat Predicted Reactivity: Pig (91%), Bovine (91%), Sheep (91%), Rabbit (82%), Dog (91%), Chicken (82%), Xenopus (82%)
Applications
Western Blot (WB), Immunohistochemistry (IHC), Peptide ELISA (EIA)
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
DB (Dot Blot) (Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.)
DB (Dot Blot) (Dot blot analysis of anti-Phospho-HIST1H3B3-S10 Phospho-specific Pab on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5ug per ml.)
WB (Western Blot) (Western blot analysis of extracts from Hela cells,untreated or treated with FBS and Calyculin A,100nM, using phospho-H3-Ps10(left) or H3 antibody(right))
WB (Western Blot) (Western blot analysis of extracts from Hela cells,untreated or treated with FBS and Calyculin A,100nM, using phospho-HIST1H3B3-S10(left) or HIST1H3B3 antibody(right))
WB (Western Blot) (Western blot analysis of lysate from Hela cell line, using Phospho-H3-pS10 Antibody. AAA28670 was diluted at 1:1000. A goat anti-rabbit IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysate at 20ug.)
WB (Western Blot) (Western blot analysis of lysates from Hela cell line, untreated or treated with FBS+calyculin A, 100ng/ml, using Phospho-H3-Ps10 Antibody, (upper) or Beta-actin (lower).)
WB (Western Blot) (Western blot analysis of lysates from Hela cell line, untreated or treated with Calyculin A, 100ng/ml, using Phospho-H3-pS10 Antibody (upper) or Beta-actin (lower).)
Protein phosphorylation is a process where a phosphate group is added to certain amino acid residues of a protein – usually serine (S), threonine (T), or tyrosine (Y) - by enzymes called kinases. This process is integral in controlling cellular signaling, cellular growth, and other biological functions.
Our catalog includes a wide range of phospho-specific antibodies that can accurately detect this important marker. They perform strongly in widely-used laboratory applications such as Western blot, flow cytometry, immunohistochemistry, and immunofluorescence microscopy. We value your trust in us and are committed to providing top-quality products and services. All of our antibodies are guaranteed to work for the applications and species indicated on our website & associated product pages.
What Are The Key Applications of Phospho Antibodies?
1. Western Blotting
One of the first steps a researcher can take in utilizing these phospho-specific antibodies, is to check if the antibody works using a technique referred to as “Western blot”. For those unfamiliar, Western Blot aids in showing whether the protein that the antibody recognizes is appearing at the correct/expected size. These phospho-specific antibodies should also be able to detect changes in the target protein’s phosphorylation (on/off state) when cells are stimulated in certain ways.
2. Staining of Fixed Cells (Immunocytochemistry)
Another routine use of these phospho-specific antibodies, is to test if the antibody is able to demonstrate similar performance when used on fixed cells (intact cells that have been preserved) as it did in the Western blot tests. It is an important aspect in many cases to confirm that the antibody works in actual intact cell samples. Ideally, the method used for cellular fixation should be the same as what is used in pathology labs (like using 10% formalin). To check if the antibody works well in tissue sections (FFPE), researchers will often test it on fixed cells that are processed similar to tissue samples.
3. Specificity Tests Using Peptides
In order to make sure that the antibody is only binding to the right target:
Laboratory technicians will mix the antibody with phospho-peptides (short segments of the protein containing the phosphate group modification).
If the antibody signal disappears, it is confirmation that it is binding to the correct phosphorylated location.
A more robust test is to use both the phosphorylated and non-phosphorylated (dephosphorylated) versions of the protein. The antibody should react only with the phosphorylated one.
Another method sometimes utilized is to treat the sample with an enzyme, such as alkaline phosphatase, that specifically removes phosphate groups. If the antibody signal disappears after this, it also confirms specificity.
4. Genetic Confirmation
As a final step, scientists can genetically manipulate the nucleotide sequence and alter the target protein by removing the exact site where phosphorylation happens. If the antibody no longer appears to detect the modified protein, it is strong evidence supporting the antibody being specific for that phosphorylated site.
Why Buy Phospho Antibodies Through Us?
The production laboratory adheres to strict and consistent protocols prior to releasing any of these phospho-specific antibodies:
Standard methods and proper controls in all tests to ensure high quality.
These antibodies are tested and validated in different cell types and species.
High quality control criterion to ensure each batch is consistent, so you will obtain reliable results every time.
FAQ
1. What Are Phospho-Specific Antibodies?
Phospho-specific antibodies are made to detect proteins only when they have a phosphate group linked to a specific amino acid residue. This empowers scientists understand if a protein is "turned on" or active, based on its phosphorylation state.
2. How to Detect Phosphorylated Proteins in a Western Blot?
To find out if a protein is phosphorylated using Western blot:
Use a phospho-specific antibody that binds only to the phosphorylated form of the protein.
You can also use a “regular” antibody for the same amino acid sequence of the protein that the phospho-specific antibody is binding to (but in this case, this antibody will not bind if there is a phosphate group present) in order to compare how much of it is phosphorylated versus how much is non-phosphorylated (or “total” protein, if the “normal” antibody’s epitopes are non-phospho-site-specific).
3. How to Choose the Best Antibody?
Here are some simple tips to help you pick the right antibody:
Know your target
Match your sample characteristics
Confirm the intended use is appropriate
Check “host” and “type”
Check the “quality” of the presented data/images
Appraise whether the available validation meets your needs
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