Calponin (Basic) Antibody Staining Protocol for Immunohistochemistry


Description: Basic calponin (calponin-h1) is a 34kD protein which exhibits a high degree of homology to acidic and neutral calponins at its N-terminal region. It is an actin, tropomyosin and calmodulin binding protein thought to be involved in the regulation of smooth muscle contraction. The expression of calponin is restricted to smooth muscle cells and is a marker of the differentiated contractile phenotype of developing smooth muscle. Vascular smooth muscle cells convert to a synthetic dedifferentiated phenotype when this protein is lost. It is thought that basic calponin exerts its effect via the cortical actin cytoskeleton. Basic calponin mRNA is expressed in smooth muscle of prostate, bowel and aorta whereas neutral and acidic calponin mRNAs are expressed in non-smooth muscle tissues such as heart, placenta, lung, kidney, pancreas, spleen, testis and ovary as well as in smooth muscle-containing tissues.


Primary Antibody

Name: Calponin (Basic) Antibody

Clone: 26A11, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CALPONIN-B

Dilution: 1:25 - 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: 60min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

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:
Tissue Type: Bowel, smooth muscle
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Bowel, smooth muscle
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


There is no need to perform antigen retrieval for acetone fixed frozen sections