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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
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Name: GIP (Y-20) Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-23554 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
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Species Reactivity: Human, mouse, rat |
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Fixation: Formalin fixed, paraffin embedded sections |
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Positive Control: Pancreas (islet) |
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Negative Control: Omit primary antibody, isotype control or absorption control |
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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 |
References: