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324865 PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb

324865
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Overview

Replacement Information

Key Specifications Table

Species ReactivityHostAntibody Type
H Rb Polyclonal Antibody

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324865-10T
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      Description
      OverviewRecognizes the ~170 kDa EGFR protein phosphorylated at Tyr845 in EGF-treated A431 cells. Note: 1 T = 1 test. Sufficient for 10 Western miniblots.
      Catalogue Number324865
      Brand Family Calbiochem®
      SynonymsAnti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵)
      Application Data
      Detection of human epidermal growth factor receptor, phosphorylated on Tyr845, by immunoblotting. Samples: Whole cell lysate from A431 cells left unstimulated (lane 1) or stimulated with EGF (lanes 2-5). Primary antibody: PhosphoDetect™ Anti-Epidermal Growth Factor Receptor (pTyr845) Rabbit pAb (Cat. No. 324865) (1:1000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphotyrosine-containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
      References
      ReferencesSieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249.
      Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067.
      Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335.
      Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139
      Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.
      Product Information
      FormLiquid
      FormulationIn Dulbecco's PBS (w/o Mg2+ and Ca2+), 1 mg/ml BSA, 50% glycerol, pH 7.4.
      Positive controlA431 cells stimulated with EGF
      Preservative≤0.1% sodium azide
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Application NotesImmunoblotting (1:1000)
      Application CommentsThe immunogen sequence is 100% conserved in mouse and rat and 93% conserved in chicken EGFR, but cross-reactivity has not been tested. When diluted to 1:1000 there is sufficient antibody to perform 10 miniblots (10 T) at 10 ml per blot. Variables associated with assay conditions will dictate the proper working dilution.
      Biological Information
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Tyr⁸⁴⁵ phosphorylation site of human EGFR
      HostRabbit
      IsotypeIgG
      Species Reactivity
      • Human
      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 Dry Ice Only
      Toxicity Standard Handling
      Storage ≤ -70°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb SDS

      Title

      Safety Data Sheet (SDS) 

      PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb Certificates of Analysis

      TitleLot Number
      324865

      References

      Reference overview
      Sieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249.
      Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067.
      Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335.
      Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139
      Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.
      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.

      Revision01-August-2007 RFH
      SynonymsAnti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵)
      ApplicationImmunoblotting (1:1000)
      Application Data
      Detection of human epidermal growth factor receptor, phosphorylated on Tyr845, by immunoblotting. Samples: Whole cell lysate from A431 cells left unstimulated (lane 1) or stimulated with EGF (lanes 2-5). Primary antibody: PhosphoDetect™ Anti-Epidermal Growth Factor Receptor (pTyr845) Rabbit pAb (Cat. No. 324865) (1:1000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphotyrosine-containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
      DescriptionRabbit polyclonal antibody adsorbed against non-phosphopeptide corresponding to the immunogen phosphorylation site and a generic tyrosine phosphorylated peptide, followed by immunoaffinity chromatography. Recognizes the ~185 KDa EGFR protein phosphorylated on Tyr845.
      BackgroundEpidermal Growth Factor (EGF) is responsible for a variety of biological effects including mitogenesis and differentiation. The EGF receptor (EGFR) has three domains: the extracellular, transmembrane, and cytoplasmic domain. Upon the binding of EGF to the extracellular domain, dimerization of the receptor occurs and the cytoplasmic domain displays tyrosine kinase activity. This leads to the autophosphorylation of the ~170 kDa glycoprotein on various residues within the cytoplasmic domain as well as the phosphorylation of intracellular substrates. Tyr845 is an autophosphorylation site within EGFR. Phosphorylation of tyrosine 845 is mediated by non-receptor tyrosine kinase c-Src.
      HostRabbit
      Immunogena synthetic phosphopeptide corresponding to amino acids surrounding the Tyr⁸⁴⁵ phosphorylation site of human EGFR
      IsotypeIgG
      Specieshuman
      Positive controlA431 cells stimulated with EGF
      FormLiquid
      FormulationIn Dulbecco's PBS (w/o Mg2+ and Ca2+), 1 mg/ml BSA, 50% glycerol, pH 7.4.
      Preservative≤0.1% sodium azide
      CommentsThe immunogen sequence is 100% conserved in mouse and rat and 93% conserved in chicken EGFR, but cross-reactivity has not been tested. When diluted to 1:1000 there is sufficient antibody to perform 10 miniblots (10 T) at 10 ml per blot. Variables associated with assay conditions will dictate the proper working dilution.
      Storage ≤ -70°C
      Avoid freeze/thaw
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
      Toxicity Standard Handling
      ReferencesSieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249.
      Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067.
      Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335.
      Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139
      Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.

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