Anti-SRP54 Antibody Picoband®

Western blot analysis of SRP54 using anti-SRP54 antibody (A06189-1).
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 placenta tissue lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human HEK293 whole cell lysates,
Lane 5: rat stomach tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse NIH/3T3 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-SRP54 antigen affinity purified polyclonal antibody (Catalog # A06189-1) at 0.25 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 SRP54 at approximately 256 kDa. The expected band size for SRP54 is at 256 kDa.

IF analysis of SRP54 using anti-SRP54 antibody (A06189-1).
SRP54 was detected in an immunocytochemical section of T47D 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-SRP54 Antibody (A06189-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Immunoprecipitating SRP54 in Jurkat whole cell lysate.
Western blot analysis of SRP54 using anti-SRP54 antibody (A06189-1).
Lane 1: Jurkat whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-SRP54 antibody in Jurkat whole cell lysate,
Lane 3: anti-SRP54 antibody (2μg) + Jurkat whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-SRP54 antigen affinity purified polyclonal antibody (A06189-1) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Light Chain). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for SRP54 at approximately 56 kDa. The expected band size for SRP54 is at 56 kDa.

Flow Cytometry analysis of U937 cells using anti-SRP54 antibody (A06189-1).
Overlay histogram showing U937 cells stained with A06189-1 (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-SRP54 Antibody (A06189-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.