Anti-ACTA2 Polyclonal Antibody is a Rabbit antibody targeting ACTA2. Anti-ACTA2 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Rabbit)

1. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (TMAB-00047)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 80% methanol (5 min at-20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. Blank control: NIH/3T3. Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (TMAB-00047)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Sample:
Liver (Rat) Lysate at 40 μg
Urinary bladder (Rat) Lysate at 40 μg
Stomach (Mouse) Lysate at 40 μg
A549 (Cell) Lysate at 30 μg
Primary: Anti-alpha smooth muscle Actin (TMAB-00047) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 46 kDa
4. Sample:
A549 (Human) Cell Lysate at 30 μg
NIH/3T3 (Mouse) Cell Lysate at 30 μg
Hela (Human) Cell Lysate at 30 μg
A431 (Human) Cell Lysate at 30 μg
HepG2 (Human) Cell Lysate at 30 μg
Stomach (Mouse) Lysate at 40 μg
Lung (Mouse) Lysate at 40 μg
Skeletal muscle (-) (Mouse) Lysate at 40 μg
Primary: Anti-alpha smooth muscle Actin (TMAB-00047) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
5. Paraformaldehyde-fixed, paraffin embedded Mouse Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Human Duodenum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Human Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Rat Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Polyclonal Antibody, Unconjugated (TMAB-00047) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
12. Blank control: NIH/3T3. Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (TMAB-00047)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
13. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (TMAB-00047), Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 μg/test.
14. 25 μg total protein per Lane of various lysates probed with alpha smooth muscle Actin polyclonal antibody, unconjugated (TMAB-00047) at 1:2000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

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

WB: 1:1000-5000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Actin alpha

Human

Myogenesis,Mesoderm,Actin,alpha smooth muscle Actin,Mesoderm,Arterial,Cytoskeleton,Actins,Smooth Muscle Cells