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

Name: p19 (G-19) Antibody

Clone: Goat polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-7403

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.

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: 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