LYAR Antibody / Cell growth-regulating nucleolar protein (FY12393)

Immunofluorescent staining of LYAR using anti-LYAR antibody (green) and anti-Beta Tubulin antibody (red). LYAR was detected in immunocytochemical section of human 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-LYAR antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated Goat Anti-Mouse IgG were 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 LYAR using anti-LYAR antibody. Lane 1: human whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human HEL whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: rat RH-35 whole cell lysates, Lane 7: mouse testis tissue lysates, Lane 8: 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-LYAR antibody at 0.25 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 LYAR is ~44 kDa.

Immunoprecipitation of LYAR in K562 whole cell lysate. Western blot analysis of LYAR using anti-LYAR antibody; Lane 1: K562 whole cell lysates (30ug); Lane 2: Rabbit control IgG instead of anti-LYAR antibody in K562 whole cell lysate; Lane 3: anti-LYAR antibody (2ug) + K562 whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-LYAR antibody at a dilution of 0.5 ug/ml and probed with a mouse anti-rabbit IgG-HRP secondary antibody. The signal is developed using ECL Plus Western Blotting Substrate. The expected molecular weight of LYAR is ~44 kDa.