Glucose-dependent Insulinotropic Polypeptide (GIP) Antibody Staining Protocol for Immunohistochemistry


Description: Glucose-dependent insulinotropic polypeptide (GIP) is a major physiologic factor in the augmentation of the insulin response to oral glucose. GIP is a peptide hormone that is released postprandially from the small intestine and acts in concert with glucagon-like peptide (GLP)-1 to potentiate glucoseinduced insulin secretion from the pancreatic beta-cell. GIP has been shown to increase adenylyl cyclase activity, elevate intracellular calcium levels, and stimulate a mitogen-activated protein kinase pathway in the pancreatic beta-cell. Additionally, nutrient protein provides a potent stimulus for GIP expression, an effect that occurs at the posttranslational level and may be mediated in part through the acid-stimulatory properties of protein. GIP release is demonstrated predominantly after ingestion of carbohydrate and fat and the effects of acid on GIP are consistent with a role for GIP as an enterogastrone.


Primary Antibody

Name: GIP (Y-20) Antibody

Clone: Goat polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-23554

Dilution: 1:50 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 minutes/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 ºC - 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 embedded sections

Positive Control: Pancreas (islet)

Negative Control: Omit primary antibody, isotype control or 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