Alpha-Tubulin Loading Control Polyclonal Antibody is a Rabbit antibody targeting Alpha-Tubulin Loading Control. Alpha-Tubulin Loading Control Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Sheep,Cow)

1. Blank control: RSC96 Cells (blue). Primary Antibody: Rabbit Anti-Tubulin-alpha 1 antibody (TMAB-00109), Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit Igg (orange), used under the same conditions); Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (TMAB-00109,1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice.
2. Sample:
Lane 1: Mouse NIH/3T3 cell lysates
Lane 2: Human HepG2 cell lysates
Lane 3: Human 293T cell lysates
Primary: Anti-Tubulin-alpha (TMAB-00109) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa
3. Sample:
Lane 1: Human HeLa cell lysates
Lane 2: Human Jurkat cell lysates
Lane 3: Human SH-SY5Y cell lysates
Lane 4: Human U-2 OS cell lysates
Lane 5: Human A431 cell lysates
Primary: Anti-Tubulin-alpha (TMAB-00109) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kDa
Observed band size: 52 kDa
4. Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Tubulin-alpha Polyclonal Antibody, Unconjugated (TMAB-00109) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Tubulin-alpha 1

Human

Tubulin