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c-jun Oncoprotein Antibody Staining Protocol for Immunohistochemistry
Description: c-jun is the normal cellular homologue of the transforming gene of avian sarcoma virus 17 and a member of the early-response gene family. The c-jun oncogene encodes a nuclear protein, p39, which is a major component of the transcription factor AP1 and interacts with the c-fos oncogene product forming a transacting heterodimer. c-jun oncoprotein plays an important role in the regulation of gene expression and signal transduction processes. Alterations in c-jun expression may affect the transcriptional initiation of specific target genes and as a consequence may affect normal cell growth and function.
Primary Antibody
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Name: c-jun oncoprotein Antibody |
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Clone: DK4, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-cJUN |
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Dilution: 1:40 - 1:80 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 minutes/room temperature |
Antigen Retrieval
| Device: Incubator |
| Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
| Heat/Cool Temperature: 37 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: Breast cancer |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: A large proportion of breast cancers are positive |
| Negative Control: Omit primary antibody, isotype control or 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 |
Notes:
Trypsin digestion is for paraffin sections. There is no need for frozen sections |
References: