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

Name: Insulin IHC Antibody

Clone: Mouse Monoclonal: ISL-8J

Supplier: IHC World

Catalog Number: IW-MA1052

Dilution: Ready to Use

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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