Affinity labeling of ras p21 protein and its use in the identification of ras p21 in cellular and tissue extracts
Journal Article
·
· J. Biol. Chem.; (United States)
OSTI ID:6596844
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), Journal Name: J. Biol. Chem.; (United States) Vol. 5; ISSN JBCHA
- Country of Publication:
- United States
- Language:
- English
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· Proceedings of the National Academy of Sciences of the United States of America; (USA)
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OSTI ID:6957947
Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BODY
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
GENES
GLANDS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLING
LIGHT NUCLEI
LIVER
MAMMALS
MAMMARY GLANDS
MATERIALS
MICE
NUCLEI
ODD-ODD NUCLEI
ONCOGENES
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SKIN
TISSUE EXTRACTS
TRACER TECHNIQUES
TRANSPLANTS
TUMOR CELLS
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BODY
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
GENES
GLANDS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLING
LIGHT NUCLEI
LIVER
MAMMALS
MAMMARY GLANDS
MATERIALS
MICE
NUCLEI
ODD-ODD NUCLEI
ONCOGENES
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SKIN
TISSUE EXTRACTS
TRACER TECHNIQUES
TRANSPLANTS
TUMOR CELLS
VERTEBRATES