VALIDATION IMAGES
Lane 1: Hela; Lane 2: SW480; Lane 3: HCT116; Lane 4: PC12 lysate probed with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:1000 overnight at 4°C followed by a conjugated secondary antibody for 60 minutes at 37°C.
Flow cytometric analysis of HeLa cells with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at a 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).
Flow cytometric analysis of SH-SY-5Y cells with ERK1/2 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.
IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in HeLa cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in MCF-7 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in NIH/3T3 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Pancreas lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Lymph node lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Rat Pancreas lysates; Lane 7: Rat Large intestine lysates; Lane 8: Rat Lymph node lysates; Lane 9: Human Hela cell lysates; Lane 10: Human SW480 cell lysates; Lane 11: Human MCF-7 cell lysates; Lane 12: Mouse NIH/3T3 cell lysates; Lane 13: Human A549 cell lysates; Lane 14: Human A431 cell lysates probed with ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) 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 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 ERK1/2 (3A12) Monoclonal Antibody(bs-R)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).
Tissue/cell:A549 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 (ERK1/2) monoclonal Antibody, Unconjugated (bsm-52259R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded Rat colon; 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 ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; 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 ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; 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 ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.
25 ug total protein per lane of various lysates (see on figure) probed with ERK1/2 monoclonal antibody, unconjugated (bsm-52259R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.