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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

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

 

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