HL-60 cell lysates probed with Anti-HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.


Formalin-fixed and paraffin embedded human cervical carcinoma labeled Anti-HIF-1-Alpha Polyclonal Antibody (bs-0737R), Unconjugated at 1:300, followed by conjugation to the secondary antibody and DAB staining


Mouse spleen cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HIF-1 Alpha Antibody(bs-0737R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


The HepG2 (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-HIF-1 Alpha antibody (bs-0737R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 μg/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.


4% Paraformaldehyde-fixed HepG2 (treated with 500uM CoCl2 for 6 hours）(H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, unconjugated (bs-0737R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.