Rabbit anti-Human, Rat NEDD4 Polyclonal Antibody | anti-NEDD4 antibody
Anti-NEDD4 Antibody Picoband
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
IHC-P: 2-5 ug/ml, Human, Rat
ICC/IF: 5 ug/ml, Human
FC/FACS: 1-3 ug/1x10^6 cells, Human
Direct ELISA: 0.1-0.5 ug/ml, Human
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P) and ICC.
FCM (Flow Cytometry)
(Figure 8. Flow Cytometry analysis of U87 cells using anti-NEDD4 antibody (AAA19413).Overlay histogram showing U87 cells stained with AAA19413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NEDD4 Antibody (AAA19413, 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 7. IF analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 was detected in an immunocytochemical section of A549 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-NEDD4 Antibody (AAA19413) 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 (Immunohistchemistry)
(Figure 6. IHC analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 was detected in a paraffin-embedded section of human renal cell carcinoma 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-NEDD4 Antibody (AAA19413) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 5. IHC analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 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-NEDD4 Antibody (AAA19413) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 was detected in a paraffin-embedded section of human liver 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-NEDD4 Antibody (AAA19413) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 3. IHC analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 was detected in a paraffin-embedded section of human colon adenocarcinoma 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-NEDD4 Antibody (AAA19413) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 2. IHC analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).NEDD4 was detected in a paraffin-embedded section of rat brain 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-NEDD4 Antibody (AAA19413) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)
WB (Western Blot)
(Figure 1. Western blot analysis of NEDD4 using anti-NEDD4 antibody (AAA19413).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 U251 whole cell lysates,Lane 2: human RT4 whole cell lysates,Lane 3: human Hacat whole cell 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-NEDD4 antigen affinity purified polyclonal antibody (#AAA19413) at 0.25 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 NEDD4 at approximately 115 kDa. The expected band size for NEDD4 is at 115 kDa.)
