Actin (alpha Smooth Muscle) Antibody Staining Protocol for Immunohistochemistry


Description: The antibody (also known as anti-alpha-Sm-1) is specific for the single isoform of alpha-smooth muscle actin. It reacts specifically witha-smooth muscle actin in immunoblotting assays and labels smooth muscle cells in frozen or formalin-fixed, paraffin-embedded tissue sections.


Primary Antibody

Name: Mouse Anti-Actin, alpha Smooth Muscle

Clone: 1A4, Mouse monoclonal

Supplier: Sigma

Catalog Number: A2547

Dilution: 1:1000 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: Antigen retrieval is not needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane/Cytoplasmic
Images: Search image

Additional Information:
Species Activity: Human, mouse, rat, bovine, chicken, frog, goat, guinea pig, rabbit, dog, sheep, snake.
Tissue Type: Smooth muscle, colon, skin
Fixation: Formalin-fixed paraffin sections or acetone fixed frozen sections as well as cell smears.
Positive Control: Smooth muscle, colon, small intestine, tongue, skin
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary