VALIDATION IMAGES
Formalin-fixed and paraffin embedded rat brain tissue labeled with Anti-BrdU(A7) Monoclonal Antibody, FITC Conjugated (bsm-0917M-FITC) at 1:200 for 40 minutes at 37°C followed by labeling Rabbit Anti-Tubulin Beta, Cy3 conjugated(bs-4511R-Cy3) 1:200, 40 minutes at 37°C
Lane 1: rat brain lysates Lane 2: rat heart lysates probed with Anti tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted band 50kD. Observed band size: 50kD.
Lane 1:Mouse heart lysates; Lane 2: Human MCF7 cell lysates probed with Rabbit Anti-Tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:300 overnight at 4˚C. Followed by a conjugated secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.
Formalin-fixed and paraffin embedded rat brain labeled with Anti-tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Mouse ovary lysates probed with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Cerebrum lysates probed with tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Uterus lysates probed with tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
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 tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, DAB staining.
Lane 1: SiHa cell lysates; Lane 2: Rat Cerebrum lysates; Lane 3: Mouse Cerebrum lysates; Lane 4: Rat Heart lysates ; Lane 5: Mouse Heart lysates probed with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Hela cell lysates; Lane 2: Mouse Cerebellum lysates; Lane 3: NIH/3T3 cell lysates; Lane 4: Mouse Cerebrum lysates ; Lane 5: HL60 cell lysates; Lane 6: Rat Cerebrum lysates; Lane 7: Rat Cerebellum lysates; Lane 8: Mouse Heart lysates probed with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
HL-60 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Beta tubulin Polyclonal Antibody(bs-4511R)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).
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 tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; 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 tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, DAB staining.
Tissue/cell:Sh-sy5y 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 (Beta tubulin) polyclonal Antibody, Unconjugated (bs-4511R) 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.
Lane 1: Mouse Cerebellum lysates; Lane 2: Rat Cerebellum lysates; Lane 3: Mouse Cerebrum lysates; Lane 4: Human HL-60 cell lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Mouse Heart lysates; Lane 7: Rat Heart lysates probed with tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Cerebrum (Mouse) Lysate at 40 ug
Primary:
Anti-Beta tubulin (bs-4511R) at 1/1000~20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Sample:
Cerebrum (Rat) Lysate at 40 ug
Primary:
Anti-Beta tubulin (bs-4511R) at 1/1000~20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Sample:
MDA-MB-231 (Human) Lysate at 40 ug
Primary:
Anti-Beta tubulin (bs-4511R) at 1/2000~20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); 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 (tubulin Beta) Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); 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 (tubulin Beta) Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse stomach); 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; Incubation with (tubulin Beta) Polyclonal Antibody, Unconjugated (bs-4511R-3) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human kidney); 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; Incubation with (tubulin Beta) Polyclonal Antibody, Unconjugated (bs-4511R-3) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Lane 1,3,5: Mouse Cerebellum tissue lysates; Lane 2,4,6: Rat Cerebellum tissue lysates probed with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:10000~1:50000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Mouse Cerebellum tissue lysates; Lane 3: Human SH-SY5Y cell lysates; Lane 4: Human HeLa cell lysates probed with Beta tubulin (Loading Control) Polyclonal Antibody, Unconjugated (bs-4511R) at 1:2000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 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 Beta tubulin Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.