VALIDATION IMAGES
HeLa cell lysates probed with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
MCF-7 cells were stained with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at [1:200] incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.
MCF7 cells(black) 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 ERK5 (8D8) Monoclonal Antibody(bsm-52069R)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).
Paraformaldehyde-fixed and paraffin-embedded Rat testis tissue incubated with ERK5 (8D8) Monoclonal Antibody (bsm-52069R) at 1:200, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed, paraffin embedded Rat testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ERK5 (8D8) Monoclonal Antibody, Unconjugated (bsm-52069R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: Mouse Testis lysates; Lane 2: Human Hela cell lysates probed with ERK5 monoclonal Antibody, Unconjugated (bsm-52069R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: MCF-7, 2: 293T, 3: NIH/3T3, 4: PC-
12
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 88 kDa
Observed MW: 115 kDa
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: MCF-7, 2: 293T, 3: NIH/3T3, 4: PC-
12
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 88 kDa
Observed MW: 115 kDa