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p15 Antibody Staining Protocol for Immunohistochemistry
Description: The normal progression of cells through the cell cycle is under the control of the cyclin-dependent protein kinases Cdk4 and Cdk6, which are subject to inhibition by the mitotic inhibitory protein p16. A member of the p16 family has been designated p15. p15 expression is upregulated approximately 30-fold in TGF b-treated human keratinocytes, suggesting that p15 may act as an effector of TGF b-mediated cell cycle arrest. The gene encoding p15 has been mapped to chromosome 9p21 at a position adjacent to the p16 gene at a site of frequent chromosomal abnormality in human tumors. It has been suggested that p15 may function as an effector of TGF b-mediated cell cycle arrest through inhibition of Cdk4 and Cdk6 kinases.
Primary Antibody
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Name: p15 (M-20) Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-1429 |
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Dilution: 1:100 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, rat, mouse |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Lung |
| 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: