p15 Antibody Staining Protocol for Immunohistochemistry


Description: The normal progression of cells through the cell cycle is under the control of the cyclin-dependent protein kinases Cdk4 and Cdk6, which are subject to inhibition by the mitotic inhibitory protein p16. A member of the p16 family has been designated p15. p15 expression is upregulated approximately 30-fold in TGF b-treated human keratinocytes, suggesting that p15 may act as an effector of TGF b-mediated cell cycle arrest. The gene encoding p15 has been mapped to chromosome 9p21 at a position adjacent to the p16 gene at a site of frequent chromosomal abnormality in human tumors. It has been suggested that p15 may function as an effector of TGF b-mediated cell cycle arrest through inhibition of Cdk4 and Cdk6 kinases.


Primary Antibody

Name: p15 (M-20) Antibody

Clone: Goat polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-1429

Dilution: 1:100 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 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Nuclear
Images: Search image

Additional Information:
Species Reactivity: Human, 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