 |
|
p53 Antibody Staining Protocol for Immunohistochemistry
Description: Mutation of the p53 protein may represent the commonest genetic event in human malignancy. Mutation results in the overexpression and thus detectability of the protein by immunohistochemistry. p53 appears to function both as an oncoprotein, perhaps by complexing in its mutant form with wild type p53, and as a tumour suppressor gene in its normal state.
Primary Antibody
|
Name: p53 Antibody |
|
Clone: Rabbit anti-Human |
|
Supplier: Novocastra |
|
Catalog Number: NCL-p53-CM1 |
|
Dilution: 1:50 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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Nuclear |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse |
| Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections |
| Positive Control: Breast or colonic carcinomas |
| 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: