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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
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Name: Calcitonin Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-CALp |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
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There is no need to perform antigen retrieval for acetone fixed frozen sections |
References: