alpha-Smooth Muscle Actin Antibody Staining Protocol for Immunohistochemistry

 

Description: ASM-1 represents an excellent marker for myogenic soft tissue tumours and smooth muscle differentiation. This antibody reacts with many types of smooth muscle cells, such as those present in vascular walls, intestinal muscularis mucosae and propria, myometrium, stroma of various tissues, and is also positive for myoepithelial cells of various glands, notably salivary and mammary gland. Myogenic soft tissues detected by ASM-1 include leiomyosarcomas, leiomyomas, and certain stromal cells surrounding infiltrating ductal carcinoma of the breast.

 

Primary Antibody

Name: alpha-Smooth Muscle Actin (alpha-SMA) Antibody

Clone: ASM-1, Mouse anti-Human

Supplier: Chemicon

Catalog Number: CBL171

Dilution: 1:50 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: No antigen retrieval 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: Cytoplasmic

Images: Search image

Additional Information:

Species Reactivity: Bovine, Chicken, Human, Mouse, Rat

Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections

Positive Control: Skin, blood vessels, GI tract (stomach, intestine, 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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