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

Name: c-jun oncoprotein Antibody

Clone: DK4, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-cJUN

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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


Trypsin digestion is for paraffin sections. There is no need for frozen sections