Cell Signaling Technology

Product Pathways - Neuroscience

Ras-GRF1 Antibody #3322

Applications Reactivity Sensitivity MW (kDa) Source
W M Transfected Only 155 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Ras-GRF1 Antibody detects transfected levels of Ras-GRF1. This antibody does not cross-react with Ras-GRF2.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal residues of mouse Ras-GRF1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected (lane 1) or transfected with either wild-type (lanes 2 and 3) or Ala916 mutant (lanes 4 and 5) Ras-GRF1, using Ras-GRF1 Antibody. Transfected COS cells were untreated or treated forskolin as indicated.(Triple HA-tagged Ras-GRF1 expressing plasmids provided by Dr. R. Mattingly, Dept. of Pharmacology, Wayne State University, Michigan.)

Background

Ras activity is regulated by GAP (GTPase activating proteins) and GEFs (guanine nucleotide exchange factors). Ras-GRF1 (also known as CDC25Mm) is neuronal RasGEF and is regulated by heterotrimeric G proteins and calcium influx (1,2). Binding to calmodulin and phosphorylation stimulate Ras-GRF1 activity (1,2). Multiple PKA phosphorylation sites on Ras-GRF have been identified. Phosphorylation on the two major sites, Ser54 and Ser822, inhibits Ras-GRF activity (3). Carbachol (a muscarinic agonist)-induced phosphorylation on Ser916 is essential but not sufficient for maximal Ras-GRF activity (4). It has been reported that Ras-GRF1 also shows GEF activity toward Rac after phosphorylation by the tyrosine kinase Src (5).

  1. Mattingly, R.R. and Macara, I.G. (1996) Nature 382, 268-72.
  2. Farnsworth, C.L. et al. (1995) Nature 376, 524-7.
  3. Baouz, S. et al. (2001) J. Biol. Chem. 276, 1742-1749.
  4. Mattingly, R.R. (1999) J Biol Chem 274, 37379-84.
  5. Kiyono, M. et al. (2000) J. Biol. Chem. 275, 29788-29793.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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