VALIDATION IMAGES
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 37˚C.
Image kindly submitted by Edita Aksamitiene as part of the Free Sample Program. Rat brain and spleen lysates probed with Rabbit Anti-HIF-1 Alpha Polyclonal Antibody (bs-0737R) at 1:1000.
L1 mouse heart lysates, L2 mouse large intestine lysates probed with Anti HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band and observed band size: 92kD
KLN 205 cells labeled with Rabbit HIF-1 Alpha Polyclonal Antibody Polyclonal Antibody, Unconjugated (bs-0737R) followed by a FITC Conjugated Secondary Antibody.
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
HeLa cells probed with HIF-1 Alpha Polyclonal Antibody, unconjugated (bs-0737R) at 1:100 dilution for 30 minutes compared to control cells (blue) and isotype control (orange)
Lane 1: Hela cell lysates; Lane 2: A431 cell lysates; Lane 3: U251 cell lysates; Lane 4: HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Hela cells(black) were fixed with 70% ice-cold methanol overnight at 4℃,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with
HIF-1 Alpha Polyclonal 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(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Hela 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 (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 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.
Lane 1: Human U-87MG cell lysates; Lane 2: Human Hela cell lysates; Lane 3: Human A431 cell lysates; Lane 4: Human U251 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
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).
Hela 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 (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 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.
Lane 1: Human Hela cell lysates; Lane 2: Human A431 cell lysates; Lane 3: Human U251 cell lysates; Lane 4: Human HepG2 cell lysates probed with HIF-1 Alpha Polyclonal Antibody, Unconjugated (bs-0737R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.