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 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 min/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: 95-100 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: 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