Anti-Ribonuclease Inhibitor/RNH1 Antibody Picoband® (monoclonal, 4F3)

Western blot analysis of Ribonuclease Inhibitor/RNH1 using anti-Ribonuclease Inhibitor/RNH1 antibody (M04147).
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 Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: human placenta 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 mouse anti-Ribonuclease Inhibitor/RNH1 antigen affinity purified monoclonal antibody (Catalog # M04147) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Ribonuclease Inhibitor/RNH1 at approximately 45 kDa. The expected band size for Ribonuclease Inhibitor/RNH1 is at 50 kDa.

IHC analysis of Ribonuclease Inhibitor/RNH1 using anti-Ribonuclease Inhibitor/RNH1 antibody (M04147).
Ribonuclease Inhibitor/RNH1 was detected in paraffin-embedded section of human mammary 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 1μg/ml mouse anti-Ribonuclease Inhibitor/RNH1 Antibody (M04147) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of Ribonuclease Inhibitor/RNH1 using anti-Ribonuclease Inhibitor/RNH1 antibody (M04147).
Ribonuclease Inhibitor/RNH1 was detected in paraffin-embedded section of human ovarian 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 1μg/ml mouse anti-Ribonuclease Inhibitor/RNH1 Antibody (M04147) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of Ribonuclease Inhibitor/RNH1 using anti-Ribonuclease Inhibitor/RNH1 antibody (M04147).
Ribonuclease Inhibitor/RNH1 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 1μg/ml mouse anti-Ribonuclease Inhibitor/RNH1 Antibody (M04147) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IF analysis of Ribonuclease Inhibitor/RNH1 using anti-Ribonuclease Inhibitor/RNH1 antibody (M04147).
Ribonuclease Inhibitor/RNH1 was detected in immunocytochemical section of Hela 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 2μg/mL mouse anti-Ribonuclease Inhibitor/RNH1 Antibody (M04147) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of A549 cells using anti- Ribonuclease Inhibitor/RNH1 antibody (M04147).
Overlay histogram showing A549 cells stained with M04147 (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 mouse anti- Ribonuclease Inhibitor/RNH1 Antibody (M04147, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.