VALIDATION IMAGES
Formalin-fixed and paraffin-embedded rat stomach labeled with Anti-Beclin 1/ATG6 Polyclonal Antibody, Unconjugated(bs-1353R) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C.
Image provided by One World Lab validation program. U138 cells probed with Rabbit Anti-Beclin 1 Polyclonal Antibody (bs-1353R) at 1:100 for 60 minutes at room temperature followed by Goat Anti-Rabbit IgG (H+L) Alexa Fluor 488 Conjugated secondary antibody.
Image provided by One World Lab validation program. SMMC7721 cells probed with Rabbit Anti-Beclin 1 Polyclonal Antibody (bs-1353R) at 1:50 for 60 minutes at room temperature followed by Goat Anti-Rabbit IgG (H+L) Alexa Fluor 488 Conjugated secondary antibody.
Lane 1: mouse brain lysates Lane 2: mouse intestine lysates probed with Anti BECN1/Beclin 1/ATG6 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:200 in 4C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 50kD.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) 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 Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:400 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Tissue/cell: 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 (Beclin 1) polyclonal Antibody, Unconjugated (bs-1353R ) 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 Mouse Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Stomch; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Stomch; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Fundus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 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 Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 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 Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 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 Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beclin 1 Polyclonal Antibody, Unconjugated (bs-1353R) at 1:800 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
25 ug total protein per lane of various lysates (see on figure) probed with Beclin 1 polyclonal antibody, unconjugated (bs-1353R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.