ERC1 Antibody / ELKS/RAB6-interacting/CAST family member 1 (FY12024)

Immunoprecipitating ERC1 in Hela whole cell lysate. Western blot analysis of ERC1 using anti-ERC1 antibody. Lane 1: Hela whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-ERC1 antibody in Hela whole cell lysate, Lane 3: anti-ERC1 antibody (2ug) + Hela whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-ERC1 antibody at a dilution of 0.5 ug/ml and probed with a goat anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. The expected band size for ERC1 is at 128 kDa.

Western blot analysis of ERC1 using anti-ERC1 antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Hela whole cell lysates, Lane 2: human Jurkat whole cell lysates Lane 3: rat brain tissue lysates, Lane 4: mouse testis tissue lysates, Lane 5: mouse NIH/3T3 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-ERC1 antibody at 0.5 ug/ml overnight at 4oC, 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 was developed using an ECL Plus Western Blotting Substrate. The expected band size for ERC1 is at 128 kDa.

Flow Cytometry analysis of Jurkat cells using anti-ERC1 antibody. Overlay histogram showing Jurkat cells stained with (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-ERC1 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Immunofluorescent staining of ERC1 using anti-ERC1 antibody (green). ERC1 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-ERC1 antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.