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Glutathione S-Transferase pi (GSTpi) Antibody Staining Protocol for Immunohistochemistry
Description:
The glutathione S-transferases (GSTs) are a multigene
family of isoenzymes which catalyse the conjugation of glutathione
to electrophilic substrates. These enzymes are involved in the
detoxification of both endogenous and exogenous electrophiles which
can react with cellular components such as DNA. The modification of
DNA by reactive compounds can initiate carcinogenesis and the GSTs
are believed to play a role in neutralising carcinogens. The
cytosolic GST isoenzymes have been classified into four evolutionary
classes; alpha, mu, pi and theta.
Primary Antibody
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Name: GSTpi Antibody |
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Clone: Rabbit polyclonal |
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Supplier: Novocastra |
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Catalog Number: NCL-GSTpi |
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Dilution: 1:500 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: Nuclear/cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Tumor |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Tumor |
| 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 |
Notes:
No need to perform antigen retrieval on frozen sections |
References: