Anti-Prostate Specific Antigen/KLK3 Antibody Picoband®

Western blot analysis of Prostate Specific Antigen/KLK3 using anti-Prostate Specific Antigen/KLK3 antibody (A01505-3).
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 30ug of sample under reducing conditions.
Lane 1: human LNCAP whole cell lysates,
Lane 2: human LNCAP whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Prostate Specific Antigen/KLK3 antigen affinity purified polyclonal antibody (Catalog # A01505-3) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Prostate Specific Antigen/KLK3 at approximately 34KD. The expected band size for Prostate Specific Antigen/KLK3 is at 34KD.

IHC analysis of Prostate Specific Antigen/KLK3 using anti-Prostate Specific Antigen/KLK3 antibody (A01505-3).
Prostate Specific Antigen/KLK3 was detected in paraffin-embedded section of human prostatic cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Prostate Specific Antigen/KLK3 Antibody (A01505-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IF analysis of Prostate Specific Antigen/KLK3 using anti-Prostate Specific Antigen/KLK3 antibody (A01505-3).
Paraffin-embedded tumor sections were used to detect Prostate Specific Antigen (PSA/KLK3). PC-3 cells were inoculated into nude mouse to establish tumors. Tumor-bearing mice were subsequently treated with docetaxel, while control mouse remained untreated. 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 1:500 rabbit anti-Prostate Specific Antigen/KLK3 Antibody (A01505-3) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (H+L) (BA1142) was used as secondary antibody at 1:500 dilution and incubated for 45 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Prostate Specific Antigen/KLK3 using anti-Prostate Specific Antigen/KLK3 antibody (A01505-3).
Paraffin-embedded tumor sections were used to detect Prostate Specific Antigen (PSA/KLK3). PC-3 cells were inoculated into nude mouse to establish tumors. Tumor-bearing mice were subsequently treated with docetaxel, while control mouse remained untreated. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:500 rabbit anti-Prostate Specific Antigen/KLK3 Antibody (A01505-3) overnight at 4°C. Two-step IHC Detection Kit (Rabbit IgG) was used as secondary antibody and incubated for 45 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.