8-OHdG Antibody Staining Protocol for Immunohistochemistry

 

Description: 8-OHdG (8-Hydroxy-2’-deoxyguanosine) is suitable for immunohistochemistry in formalin-fixed, paraffin embedded sections. 21 analogues of 8-OHdG were examined with only two exhibiting mini cross-reactivity. 8-sulfhydrylguanosine and 8-OHG demonstrate some cross-reactivity at high concentrations (at least 2 orders of magnitude were required for positive cross-reactivity.

 

Primary Antibody

Name: Mouse Anti-8-OHdG Antibody

Clone: Mouse monoclonal

Supplier: Oxis International, Inc.

Catalog Number: 24326

Dilution: 1:20

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval

Device: Steamer

Buffer/pH value: Citrate Buffer/pH 6.0

Heat/Cool Temperature: 95-100 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: Gill's Hematoxylin or Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Nuclear

Images: Search image

Additional Information:

Species Reactivity: Human

Fixation: Formalin fixed paraffin sections

Positive Control: Skin

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

 

References: