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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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