E. coli-derived human WWOX recombinant protein (Position: M1-D245).
Konjugation:
Unconjugated
Alternative Synonym:
WW domain-containing oxidoreductase, Fragile site FRA16D oxidoreductase, Short chain dehydrogenase/reductase family 41C member 1, WWOX, FOR, SDR41C1, WOX1
Anti-WWOX Antibody. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Klonalität:
Polyclonal
Konzentration:
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Application Notes: Western blot, 0.1-0.5µg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml Immunocytochemistry/Immunofluorescence, 2µg/ml Flow Cytometry (Fixed), 1-3µg/1x106 cells ELISA, 0.1-0.5µg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
Flow Cytometry analysis of U20S cells using anti-WWOX antibody. Overlay histogram showing U20S cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-WWOX Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
WB analysis of WWOX using anti-WWOX antibody.Lane 1:human MCF-7 cell
IF analysis of WWOX using anti-WWOX antibody. WWOX was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-WWOX Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of WWOX using anti-WWOX antibody. WWOX was detected in paraffin-embedded section of human prostatic cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-WWOX Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of WWOX using anti-WWOX antibody. WWOX was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-WWOX Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of WWOX using anti-WWOX antibody. WWOX was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml
Western blot analysis of WWOX using anti-WWOX antibody. 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 COLO320 whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human 293T whole cell lysates, Lane 5: human K562 whole cell lysates, Lane 6: human A549 whole cell lysates, Lane 7: human PC-3 whole cell lysates, Lane 8: human Hela 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-WWOX antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 WWOX at approximately 47 kDa. The expected band size for WWOX is at 47 kDa.
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