Calponin Antibody Staining Protocol for Immunohistochemistry


Description: Calponin, a calmodulin, binds tropomyosin and F-actin and is 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. Two isoforms of calponin exist whose molecular weights are 34kD and 29kD. Expression of the 29kD form is primarily restricted to muscle of the urogenital tract. The 34kD isoform is expressed in vascular and visceral smooth muscle of trachea, jejunum and oesophagus.


Primary Antibody

Name: Calponin Antibody

Clone: CALP, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CALPONIN

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: 60min/room temperature

Antigen Retrieval
Device: Incubator
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 37 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: Myometrium
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Myometrium
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