PCNA Antibody Staining Protocol for Immunohistochemistry

 

Description: Proliferating cell nuclear antigen (PCNA) was originally identified by immunofluorescence as a nuclear protein whose appearance correlated with the proliferative state of the cell. PCNA /cyclin has been localized by in situ hybridization to the short arm of human chromosome 20 with a peak of grains over band 20p13. PCNA gene is present in single copy and has 6 exons. It spans 4,961 bp. Synthesis of the nuclear protein cyclin  and DNA in quiescent mouse fibroblasts is coordinately induced by serum and purified growth factors. PCNA controls establishment of sister chromatid cohesion during S phase.

 

Primary Antibody

Name: PCNA IHC Antibody

Clone: Mouse Monoclonal: IML-83

Supplier: IHC World

Catalog Number: IW-MA1083

Dilution: Ready to Use

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, mouse, rat.

Fixation: Formalin fixed paraffin embedded sections or acetone fixed frozen sections and cultured cells.

Positive Control: Skin, tumor.

Negative Control: Omit primary antibody, isotype control, absorption control

Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

 

References: