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p63 Antibody Staining Protocol for Immunohistochemistry
Description: The p53 gene is a widely studied anti-oncogene, or tumor suppressor gene. The p53 gene product can act as a negative regulator of cell growth in response to DNA damage. p73 shares a high degree of homology with p53, and appears to have similar growth inhibiting and apoptosis-promoting functions. However, unlike p53, the expression of p73 is not upregulated in response to DNA damage. p73 can, when overproduced, activate the p53-responsive gene p21. p63 has also been identified based on its similarities with p53. The p63 gene encodes multiple isotypes with variable functions. p63α (also designated p51B or KET), p63β and p63γ (also designated p51A), as well as corresponding TA*p63 isoforms, contain transactivation domains which have been shown to transactivate p53 reporter genes and induce apoptosis. ΔNp63 isoforms lack the transactivation domain and can act as dominant-negative reagents to inhibit transactivation by p53 and p63.
Primary Antibody
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Name: p63 Antibody |
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Clone: Mouse monoclonal ant-Human |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-25268 |
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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. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: N/A |
| Buffer/pH value: N/A |
| Heat/Cool Temperature: N/A |
| Heat/Cool Time: N/A |
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, rat |
| Fixation: Acetone fixed frozen sections |
| Positive Control: Skin, spleen |
| 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: