TRIM21/SS-A Antibody 抗体，orb654375，Biorbyt

Flow Cytometry analysis of SiHa cells using anti-TRIM21/SS-A antibody (orb654375). Overlay histogram showing SiHa cells stained with orb654375 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIM21/SS-A Antibody (orb654375, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IF analysis of TRIM21/SS-A using anti-TRIM21/SS-A antibody (orb654375). TRIM21/SS-A was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-TRIM21/SS-A Antibody (orb654375) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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.

Western blot analysis of TRIM21/SS-A using anti-TRIM21/SS-A antibody (orb654375). 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 50ug of sample under reducing conditions. Lane 1: rat lung tissue lysates, Lane 2: mouse lung tissue lysates, Lane 3: rat thymus tissue lysates, Lane 4: mouse thymus tissue lysates, Lane 5: human HeLa whole cell lysates, Lane 6: human THP-1 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-TRIM21/SS-A antigen affinity purified polyclonal antibody (Catalog # orb654375) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for TRIM21/SS-A at approximately 52KD. The expected band size for TRIM21/SS-A is at 52KD.