Product Pathways - Neuroscience
Phospho-Ras-GRF1 (Ser916) Antibody #3321
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W | M R | Transfected Only | 155 | Rabbit |
Applications Key:
W=Western Blotting
Reactivity Key:
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-Ras-GRF1 (Ser916) Antibody detects transfected levels of Ras-GRF1 only when phosphorylated at serine 916. This antibody does not cross-react with phosphorylated Ras-GRF2.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding serine 916 of mouse Ras-GRF1. Antibodies are purified by protein A and peptide affinity chromatography.
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 Phospho-Ras-GRF1 (Ser916) Antibody (upper) or Ras-GRF1 Antibody #3322 (lower). 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).
- Mattingly, R.R. and Macara, I.G. (1996) Nature 382, 268-272.
- Farnsworth, C. L. et al. (1995) Nature 376, 524-527.
- Baouz, S. et al. (2001) J. Biol. Chem. 276, 1742-1749.
- Mattingly, R.R. (1999) J. Biol. Chem. 274, 37379-37384.
- Kiyono, M. et al. (2000) J. Biol. Chem. 275, 29788-29793.
Application References
- Yang, H. B. et al. (2003) Phosphorylation of the Ras-GRF1 exchange factor at serine-916/898 reveals activation of Ras signaling in the cerebral cortex. J. Biol. Chem. 278 (15), 13278-13285. This article references the use of Phospho-Ras-GRF1 (Ser916) Antibody in the following applications: IC-IF Western Blotting
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Companion Products
- 3322 Ras-GRF1 Antibody
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- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
