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Insulin Antibody Staining Protocol for Immunohistochemistry
Description: Insulin, synthesized by the beta cells of the islets of Langerhans, consists of 2 dissimilar polypeptide chains, A and B, which are linked by 2 disulfide bonds. The insulin gene contains 3 exons and 2 introns; exon 2 encodes the signal peptide, the B chain, and part of the C peptide, while exon 3 encodes the remainder of the C peptide and the A chain. Localization of the human insulin gene to the distal end of the short arm of chromosome 11. Harper et al. (1981)and Harper and Saunders (1981) assigned the insulin gene to 11p15.5 by in situ hybridization.
Primary Antibody
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Name: Insulin IHC Antibody |
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Clone: Mouse Monoclonal: ISL-8J |
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Supplier: IHC World |
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Catalog Number: IW-MA1052 |
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Dilution: Ready to Use |
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Incubation Time/Temp: 60 min/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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse, rat |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Pancreas |
| Negative Control: Omit primary antibody, isotype control, absorption control |
| Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
References: