553009 PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb

553009
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Overview

Replacement Information

Key Specifications Table

Species ReactivityHostAntibody Type
H, M, R Rb Polyclonal Antibody

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553009-10T
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      Description
      OverviewRecognizes c-Raf phosphorylated at Tyr340 and Tyr341 in HEK293 cells overexpressing c-Raf.
      Note: 1 T = 1 test. Sufficient for 10 Western miniblots.
      Catalogue Number553009
      Brand Family Calbiochem®
      Application Data
      Detection of human c-Raf, phosphorylated on Tyr340, by immunoblotting. Samples: Whole cell lysate from HEK293 cells overexpressing c-Raf left unstimulated (lane 1) or stimulated with active Src (lane 2-5). Primary antibody: PhosphoDetect™ Anti-c-Raf (pTyr340, Tyr341) Rabbit pAb (Cat. No. 553009) (1:000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphoserine containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
      References
      ReferencesYan, F., and Polk, D.B. 2001. Cancer Res. 61, 963.
      Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151.
      Mason, C.S. et al. 1999. EMBO J. 18, 2137.
      Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400.
      Moelling, K. et al. 1984. Nature 312, 558.
      Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.
      Product Information
      FormLiquid
      FormulationIn phosphate buffer (without Mg2+ and Ca2+), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).
      Positive controlHEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunocytochemistry
      Application NotesImmunoblotting (1:1000)
      Immunocytochemistry (see comments)
      Application CommentsThe immunogen sequence is 100% conserved in mouse and rat, but cross-reactivity has not been tested. This antibody has also been reported to work for immunocytochemistry. Variables associated with assay conditions will dictate the proper working dilution.
      Biological Information
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)
      ImmunogenHuman
      HostRabbit
      IsotypeIgG
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Antibody TypePolyclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb SDS

      Title

      Safety Data Sheet (SDS) 

      PhosphoDetect™ Anti-c-Raf (pTyr³⁴⁰, Tyr³⁴¹) Rabbit pAb Certificates of Analysis

      TitleLot Number
      553009

      References

      Reference overview
      Yan, F., and Polk, D.B. 2001. Cancer Res. 61, 963.
      Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151.
      Mason, C.S. et al. 1999. EMBO J. 18, 2137.
      Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400.
      Moelling, K. et al. 1984. Nature 312, 558.
      Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.

      Brochure

      Title
      MAPK Pathway Poster ( 750 KB )
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision15-August-2007 RFH
      ApplicationImmunoblotting (1:1000)
      Immunocytochemistry (see comments)
      Application Data
      Detection of human c-Raf, phosphorylated on Tyr340, by immunoblotting. Samples: Whole cell lysate from HEK293 cells overexpressing c-Raf left unstimulated (lane 1) or stimulated with active Src (lane 2-5). Primary antibody: PhosphoDetect™ Anti-c-Raf (pTyr340, Tyr341) Rabbit pAb (Cat. No. 553009) (1:000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphoserine containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
      DescriptionRabbit polyclonal antibody adsorbed against non-phosphopeptide corresponding to the immunogen phosphorylation site, followed by immunoaffinity chromatography. Recognizes the ~74 KDa c-Raf protein phosphorylated on Tyr340/341.
      BackgroundThe raf family of proto-oncogenes consists of three members: A-raf-1, B-raf, and c-raf-1 which show the greatest degree of homology in the carboxy-terminal half of the molecule. In the case of c-raf-1 this region has been shown to have protein (serine/threonine) kinase activity. C-raf was first identified in an oncogeneic form, v-raf, which is the oncogene of the transforming virus MSV. The raf-1 gene is the most studied of the three and encodes a protein with an apparent molecular mass of ~74 kDa. Raf is widely expressed and plays an important role in cell proliferation and differentiation. Raf relays signals related to the activation of growth factor receptor to the activation of MAPK and utlimately to the nucleus. Activation of raf itself is dependent on tyrosine (341) phosphorylation. Tyrosine phosphorylation is accomplished by Src tyrosine kinase.
      HostRabbit
      Immunogen speciesHuman
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Tyr³⁴⁰ and Tyr³⁴¹, phosphorylation sites of human c-Raf (Raf-1)
      IsotypeIgG
      Specieshuman, mouse, rat
      Positive controlHEK293 cells over expressing c-Raf, phosphorylated in vitro by Src.
      FormLiquid
      FormulationIn phosphate buffer (without Mg2+ and Ca2+), 1 mg/ml BSA, 50% glycerol, pH 7.3 (+/- 0.1).
      Preservative≤0.1% sodium azide
      CommentsThe immunogen sequence is 100% conserved in mouse and rat, but cross-reactivity has not been tested. This antibody has also been reported to work for immunocytochemistry. Variables associated with assay conditions will dictate the proper working dilution.
      Storage -20°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Standard Handling
      ReferencesYan, F., and Polk, D.B. 2001. Cancer Res. 61, 963.
      Yip-Schneider, M.T., et al. 2000. Biochem. J. 351, 151.
      Mason, C.S. et al. 1999. EMBO J. 18, 2137.
      Bonner, T.I. et al. 1985. Mol. Cell. Biol. 5, 1400.
      Moelling, K. et al. 1984. Nature 312, 558.
      Rapp, U.R. et al. 1983. Proc. Natl. Acad. Sci. USA 80, 4218.

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