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Cdk2 Antibody Staining Protocol for Immunohistochemistry
Description: In vertebrates, as in yeast, multiple cyclins have been identified, including a total of eight such regulatory proteins in mammals. In contrast to the situation in yeast, the Cdc2 p34 kinase is not the only catalytic subunit identified in vertebrates that can interact with cyclins. While Cdc2 p34 is essential for the G2 to M transition in vertebrate cells, a second Cdc2-related kinase has also been implicated in cell cycle control. This protein designated cyclin dependent kinase 2 (Cdk2) p33, also binds to cyclins and its kinase activity is temporally regulated during the cell cycle. Several additional Cdc2 p34-related cyclin dependent kinases have been identified. These include Cdk3, Cdk4, Cdk5, PCTAIRE-1, PCTAIRE-2, PCTAIRE-3, Cdk6, Cdk7 and Cdk8.
Primary Antibody
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Name: Cdk2 Antibody |
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Clone: Rabbit anti-Cdk2 |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-163 |
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Dilution: 1:3000 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:
| Tissue Type: Colon |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Colon |
| 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: