Preparation and Storage
推奨アッセイ手順
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml .
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Raf (c-Raf-1) is a cytoplasmic serine/threonine protein kinase and is a member of a family of proteins which are highly conserved from Drosophila to mammals. Raf has a critical role in the response to many growth factors including: EGF, PDGF, insulin, IL-2, IL-3, CSF-1, and GM-CSF. Raf can directly interact with Ras-GTP and subsequently become activated. Thus Raf plays a prominent role in the Ras signaling pathway by transferring a signal from Ras which has been activated by growth factor receptor-stimulated tyrosine kinase activity. This leads to the stimulation and activation of a number of other cytoplasmic serine/threonine kinases. Raf regulates the MAP kinase pathway by phosphorylating and activating MEK, which then phosphorylates and activates MAP kinase (ERK). ERK then phosphorylates and activates Rsk. Raf activity can also be regulated independently of Ras. Cyclic AMP (cAMP) activation of protein kinase A (PKA) can inhibit growth factor stimulation of Raf.
Development References (5)
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Bondzi C, Grant S, Krystal GW. A novel assay for the measurement of Raf-1 kinase activity. Oncogene. 2000; 19(43):5030-5033. (Clone-specific: Immunoprecipitation, In vitro kinase assay, Western blot). View Reference
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Cheng JJ, Wung BS, Chao YJ, Wang DL. Sequential activation of protein kinase C (PKC)-alpha and PKC-epsilon contributes to sustained Raf/ERK1/2 activation in endothelial cells under mechanical strain. J Biol Chem. 2001; 276(33):31368-31375. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Coles LC, Shaw PE. PAK1 primes MEK1 for phosphorylation by Raf-1 kinase during cross-cascade activation of the ERK pathway. Oncogene. 2002; 21(14):2236-2244. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Rapp UR. Role of Raf-1 serine/threonine protein kinase in growth factor signal transduction. Oncogene. 1991; 6(4):495-500. (Biology). View Reference
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Rizzo MA, Kraft CA, Watkins SC, Levitan ES, Romero G. Agonist-dependent traffic of raft-associated Ras and Raf-1 is required for activation of the mitogen-activated protein kinase cascade. J Immunol. 2001; 276(37):34928-34933. (Clone-specific: Immunofluorescence, Western blot). View Reference
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