VALIDATION IMAGES
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 Phospho-eIF-2a(S51) Antibody, Unconjugated (bsm-54490R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse pancreas; 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 Phospho-eIF-2a(S51) Antibody, Unconjugated (bsm-54490R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human liver; 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 Phospho-eIF-2a(S51) Antibody, Unconjugated (bsm-54490R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human prostate cancer; 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 Phospho-eIF-2a(S51) Antibody, Unconjugated (bsm-54490R) at 1:50 for 30 minutes at room temperature, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human pancreas; 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 Phospho-eIF-2a(S51) Antibody, Unconjugated (bsm-54490R) at 1:50 for 30 minutes at room temperature, DAB staining.
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 (Phospho-eIF-2a(S51)) Monoclonal Antibody, Unconjugated (bsm-54490R) 1:50, 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.
Flow cytometric analysis of Phospho-eIF-2a(S51) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (bsm-54490R, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).