Actin (alpha Smooth Muscle) Antibody Staining Protocol for Immunohistochemistry

 

Description: This antibody reacts with the alpha-smooth muscle isoform of actin present in smooth muscle cells of vessels and different parenchymes, myoepithelial cells, pericytes and some stromal cells in the intestine, testis and ovary. It does not react with actin from fibroblasts (beta-and gamma-cytoplasmic), striated muscle (alpha-sarcomic), and myocardium (alpha-myocardial).

 

Primary Antibody

Name: Mouse Anti-Human Actin, alpha Smooth Muscle

Clone: 1A4

Supplier: Dako

Catalog Number: M0851

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.

Fixation: Formalin-fixed paraffin sections or acetone fixed frozen sections as well as cell smears.

Positive Control: Smooth muscle, colon

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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