RBL2 Antibody

Immunohistochemical staining of mouse tissue using anti-pRb2/p130 antiserum. The staining shows the location of pRb2/p130 in developing mouse tissue. Other detection systems should yield similar results. Sections were cut at 5-7 µm, mounted on glass and dried overnight at 37°C. All sections were deparaffinized in xylene, rehydrated through a graded alcohol series and washed in phosphate-buffered saline (PBS). PBS was used for all subsequent washes and for antiserum dilution. Tissue sections were quenched sequentially in 0.5% hydrogen peroxide and blocked with diluted 10% normal goat anti-rabbit serum. Slides were incubated at 20°C for 1 h with rabbit anti-pRb2/p130 (1:500) dilution, washed, and then reacted with diluted goat anti-rabbit biotinylated antibody for 30 min. Slides were then reacted with streptavidin-peroxidase conjugate for 30 min at 20°C. Diaminobenzidine was used as the final chromogen. Negative controls for each tissue section were prepared by substituting the primary antiserum with pre-immune serum.

Western Blot of Rabbit Anti-Rb2 p130 Antibody. Lane 1: HEK 293 pcDNA3. Lane 2: HEK 293 pcDNA3-Rb2wt. Lane 3: HEK 293 pcDNA3-Rb2-PM19. Load: 30 µg per lane. Primary antibody: Anti-Rb2 antibody at 1:250 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 130 kDa for p130/Rb2.