Affinity labeling of ras p21 protein and its use in the identification of ras p21 in cellular and tissue extracts

Journal Article · Sun Feb 15 00:00:00 EST 1987 · J. Biol. Chem.; (United States)

OSTI ID:6596844

Basu, A; Modak, M J

We have carried out photoaffinity labeling of the ras p21 protein, a ras oncogene product, with (alpha-/sup 32/P)GTP. Based on our studies, a sensitive, rapid, and specific assay for the detection of multiple forms of ras p21 has been developed. The specificity of this protocol is shown by (a) sensitivity of affinity labeling of ras p21 to known inhibitors of GTP binding and (b) immunoprecipitation of affinity labeled protein with anti-ras p21 serum. Detection and semiquantitation of ras p21 by this method is accomplished in less than 24 h and requires as little as 100,000 cells or about 5 mg of tissue sample from skin tumor, liver, and mammary tumor tissues. Furthermore, using this approach, we were able to detect the selective loss of one species of ras p21 in transplanted Morris hepatoma cells.

• Research Organization: Univ. of Medicine and Dentistry of New Jersey, Newark
• OSTI ID: 6596844
• Journal Information: J. Biol. Chem.; (United States), Vol. 5
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
BIOCHEMICAL REACTION KINETICS
LABELLING
TUMOR CELLS
TRANSPLANTS
AFFINITY
LIVER
MAMMARY GLANDS
MICE
ONCOGENES
PHOSPHORUS 32
SKIN
TISSUE EXTRACTS
TRACER TECHNIQUES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BODY
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
GENES
GLANDS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MATERIALS
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
REACTION KINETICS
RODENTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques