VALIDATION IMAGES
Lane 1: Mouse Macrophages probed with Rabbit Anti-HMGB1 Polyclonal Antibody, Unconjugated (bs-0664R) 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 endometrium carcinoma labeled Anti-HMGB1 Polyclonal Antibody, Unconjugated (bs-0664R) at 1:200, followed by conjugation to the secondary antibody and DAB staining
Human HepG2 cell lysates probed with Rabbit Anti-HMGB1 Polyclonal Antibody, PE-Cy5 conjugated (bs-0664R-PE-Cy5) (green) at 1:20 for 30 minutes followed compared to unstained cells (blue) and PE-Cy5 isotype control bs-0295P-PE-Cy5(orange).
Rat splenocytes stained with Anti- HMGB1 Polyclonal Antibody, A488 Conjugated (bs-0664R-A488) at 1:50.
HL-60 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 HMGB1 Polyclonal Antibody(bs-0664R)at 1:100 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).
HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (HMGB1) polyclonal Antibody, Unconjugated (bs-0664R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.