Antibody Type: Mouse Monoclonal

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

Reactivity: Human,Mouse,Rat (predicted:Chicken)

IgG

6K554

Human,Mouse,Rat (predicted:Chicken)

1. Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of TMAB-00045) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructions and DAB staining.
2. Blank control: NIH/3T3. Primary Antibody (green line): Mouse Anti-alpha smooth muscle Actin antibody (TMAB-00045)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-mouse IgG-FITC
Dilution: 0.5 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. Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (rat uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
10. Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
11. 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 Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
12. 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 Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
13. 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 Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
14. Paraformaldehyde-fixed, paraffin embedded Human Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
15. Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with alpha smooth muscle Actin Monoclonal Antibody, Unconjugated (TMAB-00045) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Mouse IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
16. 25 μg total protein per Lane of various lysates probed with alpha smooth muscle Actin monoclonal antibody, unconjugated (TMAB-00045) at 1:1000 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: 1:500-5000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/Test

Monoclonal

KLH conjugated synthetic peptide: human Actin alpha

Human

59

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