Hela (H) cells were treated with or without thapsigargin (300 nM) for 30 min, 25 μg total protein per lane of cell lysates (see on figure) probed with Phospho-EIF2S1 (Ser51) monoclonal antibody, unconjugated (bsm-52306R) 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 Human Pancreatic Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Mouse Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Phospho-EIF2S1 (Ser51) Monoclonal Antibody, Unconjugated(bsm-52306R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-EIF2S1 (S51) antibody (bsm-52306R) at 1/200 dilution. A: Untreated human breast carcinoma tissue B: λ-PPase treated human breast carcinoma tissue C: Negative control The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52306R) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.