Amylin Peptide Antibody Staining Protocol for Immunohistochemistry

 

Description: Amylin is a pancreatic islet peptide with a role in the maintenance of glucose homeostasis. It is predominantly found in the beta cells of the pancreas and to a lesser extent in the gastrointestinal tract and nervous system. Amylin appears to work with insulin to regulate plasma/glucose concentrations in the bloodstream. Two fragments of amylin have been identified in vivo. One fragment contains residues 17 to 37 of human amylin and the other contains residues 24 to 37. Amylin (24-37) peptide fragment forms amyloid deposits, however, smaller fragments eg residues 20 to 29, are also capable of generating amyloid.

 

Primary Antibody

Name: Amylin Peptide Antibody

Clone: R10/99, Mouse

Supplier: Novocastra Labs

Catalog Number: NCL-AMYLIN

Dilution: 1:50 - 1:100 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: 60 min/room temperature

Antigen Retrieval

Device: Antigen retrieval not needed

Buffer/pH value: N/A

Heat/Cool Temperature: N/A

Heat/Cool Time: N/A

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 and Secreted

Images: Search image

Additional Information:

Tissue Type: Pancreas

Fixation: Formalin-fixed paraffin sections

Positive Control: Pancreas

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

 

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