LSM14A Antibody / RAP55 / AlphaSNBP (FY12475)

Immunofluorescent staining of LSM14A using anti-LSM14A antibody (green). LSM14A was detected in an immunocytochemical section of HELA 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-LSM14A 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 nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of LSM14A using anti-LSM14A antibody. Lane 1: human HEL whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: human 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-LSM14A 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 enhanced chemiluminescent. The expected molecular weight of LSM14A is ~51 kDa.

Flow Cytometry analysis of SH-SY5Y cells using anti-LSM14A antibody. Overlay histogram showing SH-SY5Y 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-LSM14A 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.

Immunoprecipitation of LSM14A in whole cell lysate. Western blot analysis of LSM14A using anti-LSM14A antibody. Lane 1: whole cell lysates (30ug) Lane 2: Rabbit control IgG instead of anti-LSM14A antibody in whole cell lysate. Lane 3: anti-LSM14A antibody (2ug) + whole cell lysate (500ug) After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-LSM14A 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 molecular weight of LSM14A is ~51 kDa.