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product-image-AAA126095_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of Raji cells using anti-ASK1/MAP3K5 antibody (AAA126095).Overlay histogram showing Raji cells stained with AAA126095 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ASK1/MAP3K5 Antibody (AAA126095, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit ASK1/MAP3K5 Polyclonal Antibody | anti-MAP3K5 antibody

Anti-ASK1/MAP3K5 Antibody Picoband

Gene Names
MAP3K5; ASK1; MEKK5; MAPKKK5
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunocytochemistry, Immunohistochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
ASK1/MAP3K5, Antibody; Anti-ASK1/MAP3K5 Antibody Picoband; anti-MAP3K5 antibody
Ordering
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-MAP3K5 antibody
WB (Western Blot), ICC (Immunocytochemistry), IHC (Immunohistochemistry), IF (Immunofluorescence), FCM/FACS (Flow Cytometry), ELISA
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human ASK1/MAP3K5 recombinant protein (Position: A178-K1112).
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM/FACS (Flow Cytometry)

(Figure 5. Flow Cytometry analysis of Raji cells using anti-ASK1/MAP3K5 antibody (AAA126095).Overlay histogram showing Raji cells stained with AAA126095 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ASK1/MAP3K5 Antibody (AAA126095, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA126095_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of Raji cells using anti-ASK1/MAP3K5 antibody (AAA126095).Overlay histogram showing Raji cells stained with AAA126095 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ASK1/MAP3K5 Antibody (AAA126095, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 4. IF analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

product-image-AAA126095_IF10.jpg IF (Immunofluorescence) (Figure 4. IF analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/mL rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

IHC (Immunohistochemisry)

(Figure 3. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

product-image-AAA126095_IHC11.jpg IHC (Immunohistochemisry) (Figure 3. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

product-image-AAA126095_IHC13.jpg IHC (Immunohiostchemistry) (Figure 2. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).ASK1/MAP3K5 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-ASK1/MAP3K5 Antibody (AAA126095) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human RT4 whole cell lysates,Lane 2: human HEL whole cell lysates,Lane 3: human HL-60 whole cell lysates,Lane 4: human THP-1 whole cell lysates,Lane 5: rat liver tissue lysates,Lane 6: mouse liver tissue lysates,After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ASK1/MAP3K5 antigen affinity purified polyclonal antibody (#AAA126095) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ASK1/MAP3K5 at approximately 155 kDa. The expected band size for ASK1/MAP3K5 is at 155 kDa.)

product-image-AAA126095_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (AAA126095).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.Lane 1: human RT4 whole cell lysates,Lane 2: human HEL whole cell lysates,Lane 3: human HL-60 whole cell lysates,Lane 4: human THP-1 whole cell lysates,Lane 5: rat liver tissue lysates,Lane 6: mouse liver tissue lysates,After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ASK1/MAP3K5 antigen affinity purified polyclonal antibody (#AAA126095) at 0.5 ug/mL overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ASK1/MAP3K5 at approximately 155 kDa. The expected band size for ASK1/MAP3K5 is at 155 kDa.)
Related Product Information for anti-MAP3K5 antibody
Apoptosis signal-regulating kinase 1 (ASK1) also known as mitogen-activated protein kinase 5 (MAP3K5) is a member of MAP kinase family and as such a part of mitogen-activated protein kinase pathway. Mitogen-activated protein kinase (MAPK) signaling cascades include MAPK or extracellular signal-regulated kinase (ERK), MAPK kinase (MKK or MEK), and MAPK kinase kinase (MAPKKK or MEKK). MAPKK kinase/MEKK phosphorylates and activates its downstream protein kinase, MAPK kinase/MEK, which in turn activates MAPK. The kinases of these signaling cascades are highly conserved, and homologs exist in yeast, Drosophila, and mammalian cells. MAPKKK5 contains 1,374 amino acids with all 11 kinase subdomains. Northern blot analysis shows that MAPKKK5 transcript is abundantly expressed in human heart and pancreas. The MAPKKK5 protein phosphorylates and activates MKK4 (aliases SERK1, MAPKK4) in vitro, and activates c-Jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) during transient expression in COS and 293 cells; MAPKKK5 does not activate MAPK/ERK.
Product Categories/Family for anti-MAP3K5 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
154,537 Da
NCBI Official Full Name
mitogen-activated protein kinase kinase kinase 5
NCBI Official Synonym Full Names
mitogen-activated protein kinase kinase kinase 5
NCBI Official Symbol
MAP3K5
NCBI Official Synonym Symbols
ASK1; MEKK5; MAPKKK5
NCBI Protein Information
mitogen-activated protein kinase kinase kinase 5; ASK-1; MEKK 5; MEK kinase 5; MAP/ERK kinase kinase 5; MAPK/ERK kinase kinase 5; apoptosis signal regulating kinase 1; apoptosis signal-regulating kinase 1
UniProt Protein Name
Mitogen-activated protein kinase kinase kinase 5
UniProt Gene Name
MAP3K5
UniProt Synonym Gene Names
ASK1; MAPKKK5; MEKK5; ASK-1; MEK kinase 5; MEKK 5
UniProt Entry Name
M3K5_HUMAN

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Product Notes

The MAP3K5 map3k5 (Catalog #AAA126095) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-ASK1/MAP3K5 Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's ASK1/MAP3K5 can be used in a range of immunoassay formats including, but not limited to, WB (Western Blot), ICC (Immunocytochemistry), IHC (Immunohistochemistry), IF (Immunofluorescence), FCM/FACS (Flow Cytometry), ELISA. Researchers should empirically determine the suitability of the MAP3K5 map3k5 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ASK1/MAP3K5, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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