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p19 ARF Antibody Staining Protocol for Immunohistochemistry
Description:
The progression of cells through the cell cycle is regulated by a
family of proteins designated cyclin-dependent kinases (Cdks).
Sequential activation of individual members of this family and their
consequent phosphorylation of critical substrates, promote orderly
progression through the cell cycle. The protein p16INK4A, identified
as a negative regulator of the cell cycle, has been shown to bind to
and inhibit the activity of the Cdk4/cyclin D complex. p19 ARF,
which is unrelated to p16, arises from transcription of an
alternative reading frame of the p16 gene. Like p16, p19 ARF has
been shown to induce cell cycle arrest. Mice lacking p19 ARF but
expressing functional p16 have been shown to develop tumors early in
life. Further studies have indicated that p19 ARF may be disrupted
in a large percentage of human T cell acute lymphoblastic leukemias.
Primary Antibody
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Name: p19 (G-19) Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-7403 |
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Dilution: 1:200 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: 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: