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

Name: GSTpi Antibody

Clone: Rabbit polyclonal

Supplier: Novocastra

Catalog Number: NCL-GSTpi

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.

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: