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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
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Name: Mouse Anti-8-OHdG Antibody |
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Clone: Mouse monoclonal |
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Supplier: Oxis International, Inc. |
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Catalog Number: 24326 |
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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. |
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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 |
Counterstain
| Reagent: 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: