Anti-C14orf166/RTRAF Antibody Picoband®

Western blot analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human PC-3 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human RT4 whole cell lysates,
Lane 4: human SIHA whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat heart tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse heart 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-C14orf166/RTRAF antigen affinity purified polyclonal antibody (A31823-2) at 0.5 μg/mL overnight at 4°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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for C14orf166/RTRAF at approximately 28 kDa. The expected band size for C14orf166/RTRAF is at 28 kDa.

IHC analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
C14orf166/RTRAF was detected in a paraffin-embedded section of human ovarian 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 μg/ml rabbit anti-C14orf166/RTRAF Antibody (A31823-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
C14orf166/RTRAF 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 μg/ml rabbit anti-C14orf166/RTRAF Antibody (A31823-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
C14orf166/RTRAF was detected in a paraffin-embedded section of human colon 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 μg/ml rabbit anti-C14orf166/RTRAF Antibody (A31823-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
C14orf166/RTRAF was detected in a paraffin-embedded section of human tonsil 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 μg/ml rabbit anti-C14orf166/RTRAF Antibody (A31823-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2).
C14orf166/RTRAF was detected in a paraffin-embedded section of human prostate 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 μg/ml rabbit anti-C14orf166/RTRAF Antibody (A31823-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of C14orf166/RTRAF using anti-C14orf166/RTRAF antibody (A31823-2) and anti-Beta Tubulin antibody (M01857-3).
C14orf166/RTRAF was detected in an immunocytochemical section of U2OS 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 μg/mL rabbit anti-C14orf166/RTRAF Antibody (A31823-2) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight?488 Conjugated Goat Anti-Rabbit IgG (BA1127) and DyLight?594 Conjugated Goat Anti-Mouse IgG (BA1141) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.