Calcitonin Antibody Staining Protocol for Immunohistochemistry


Description: Calcitonin is a peptide hormone synthesised by the parafollicular cells of the thyroid. It causes reduction in serum calcium, an effect opposite to that of parathyroid hormone (PTH). Human calcitonin contains 32 amino acids and has a molecular weight of 3,421kD. Multiple calcitonin polypeptides are encoded in a single messenger RNA. Evidence exists that indicates alternative RNA splicing of the transcripts of the calcitonin gene is responsible for the production of different polypeptide products. Genomic mapping results are consistent with the existence of a single calcitonin gene.


Primary Antibody

Name: Calcitonin Antibody

Clone: Rabbit anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CALp

Dilution: 1:100 - 1:200 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: C cells of thyroid
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: C cells of thyroid
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