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PK04 Raytide™ EL Substrate

Raytide™ EL Substrate : FDS (Fiches de données de sécurité), certificats d’analyse (CoA) et de qualité (CoQ), dossiers, brochures et autres documents disponibles.
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      Replacement Information
      Description
      Overview

      This product has been discontinued.





      A modified gastrin analog that serves as a general tyrosine kinase substrate. Exhibits a higher labeling efficiency than the standard Raytide™ Substrate (Cat. No. PK02).
      Catalogue NumberPK04
      Brand Family Calbiochem®
      References
      Product Information
      FormLyophilized
      FormulationLyophilized from a volatile buffer, BSA.
      Applications
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      Application CommentsMaterials Needed:
      • Protein tyrosine Kinase (such as p60c-src, Cat# PK03)
      • [γ-32P]ATP (10 mCi/ml)
      • Kinase Assay Buffer: 50 mM HEPES, pH 7.5 containing 0.1 mM EDTA and 0.015% Brij® 35 detergent
      • Kinase Dilution Buffer: Kinase assay buffer containing 0.1 mg/mL BSA and 0.2% β-mercaptoethanol (note: b-mercaptoethanol is required for src kinase activity)
      • ATP Mix: 0.15 mM ATP, 30 mM MgCl2 and 200 µCi [γ-32P]ATP per ml in kinase assay buffer
      • Phosphoric acid
      • Acetone
      • P81 Ion Exchange Chromatography paper (i.e. Whatman #3698915)

      Procedure:
      • Dilute kinase protein solution in Kinase Dilution Buffer if of sufficiently high concentration or, alternatively, buffer exchange the kinase protein of interest into the Kinase Dilution Buffer.
      • Mix 10 µl diluted kinase with 10 µl Raytide™ EL (1 mg/ml solution in Kinase Assay Buffer).
      • Start the reaction by adding 10 µl of ATP Mix.
      • Incubate at 30°C for 30 min.
      • Stop the reaction by adding 120 µl of 10% phosphoric acid. Vortex and apply 120 µl onto a square (2 cm x 2 cm) of P81 paper.
      • Wash papers extensively in 0.5% phosphoric acid. Wash once with acetone. Dry and quantitate by liquid scintillation counting. (Phosphorylated Raytide™ EL will stick, while ATP will wash off the papers).
      Biological Information
      Purity>95 by HPLC
      Physicochemical Information
      Peptide SequenceH-Lys-Lys-Lys-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Leu-Asp-Phe-OH
      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 +2°C to +8°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C) for long-term storage or refrigerate (4°C) for short-term storage with 0.1% azide. Avoid freeze/thaw cycles of solutions.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Référence GTIN
      PK04 0

      Documentation

      Raytide™ EL Substrate Certificats d'analyse

      TitreNuméro de lot
      PK04

      Brochure

      Titre
      Protein Phosphatases Technical Bulletin
      Fiche technique

      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.

      Revision20-February-2008 JSW
      DescriptionProtein Tyrosine Kinase substrate Raytide™ EL ("EL" for "enhanced labeling") is a modified gastrin analog. Raytide™ EL differs from the standard Raytide™ substrate (Cat. No. PK02); in that it provides a higher labeling efficiency when used as either a kinase or a phosphatase substrate. The carboxyl end of the peptide contains acidic and hydrophilic amino acid residues, while the amino terminus contains basic amino acids. In phosphoric acid the amino end is positively charged and binds to phosphocellulose paper (p81 Whatman). The distance between the two charged ends of the peptide prevents neutralization. Additionally, the peptide maintains an unfolded state and functions as an excellent substrate for protein tyrosine kinases (see procedure section) and tyrosine phosphates (Nature 350:359-362, 1991).
      BackgroundOncogene Research Products Protein Tyrosine Kinase substrate Raytide™ EL ("EL" for "enhanced labeling") is a modified gastrin analog. Raytide™ EL differs from the standard Raytide™ substrate (Cat. No. PK02); in that it provides a higher labeling efficiency when used as either a kinase or a phosphatase substrate. The carboxyl end of the peptide contains acidic and hydrophilic amino acid residues, while the amino terminus contains basic amino acids. In phosphoric acid the amino end is positively charged and binds to phosphocellulose paper (p81 Whatman). The distance between the two charged ends of the peptide prevents neutralization. Additionally, the peptide maintains an unfolded state and functions as an excellent substrate for protein tyrosine kinases (see procedure section) and tyrosine phosphates (Nature 350:359-362, 1991). Note: The exact sequence of Raytide™ and Raytide™ EL are proprietary to Oncogene Research Products and cannot be released.
      FormLyophilized
      FormulationLyophilized from a volatile buffer, BSA.
      Recommended reaction conditions
      Protocol for Using Raytide in a Kinase Assay Materials Needed: • Protein tyrosine Kinase (such as p60c-src, Cat# PK03) • [γ-32P]ATP (10 mCi/ml) • Kinase Assay Buffer: 50 mM HEPES, pH 7.5 containing 0.1 mM EDTA and 0.015% Brij® 35 detergent • Kinase Dilution Buffer: Kinase assay buffer containing 0.1 mg/mL BSA and 0.2% β-mercaptoethanol (note: β-mercaptoethanol is required for src kinase activity) • ATP Mix: 0.15 mM ATP, 30 mM MgCl2 and 200 µCi [γ-32P]ATP per ml in kinase assay buffer • Phosphoric acid • Acetone • P81 Ion Exchange Chromatography paper (i.e. Whatman #3698915) Procedure: • Dilute kinase protein solution in Kinase Dilution Buffer if of sufficiently high concentration or, alternatively, buffer exchange the kinase protein of interest into the Kinase Dilution Buffer. • Mix 10 µl diluted kinase with 10 µl Raytide™ EL (1 mg/ml solution in Kinase Assay Buffer). • Start the reaction by adding 10 µl of ATP Mix. • Incubate at 30°C for 30 min. • Stop the reaction by adding 120 µl of 10% phosphoric acid. Vortex and apply 120 µl onto a square (2 cm x 2 cm) of P81 paper. • Wash papers extensively in 0.5% phosphoric acid. Wash once with acetone. Dry and quantitate by liquid scintillation counting. (Phosphorylated Raytide™ EL will stick, while ATP will wash off the papers).
      Peptide SequenceH-Lys-Lys-Lys-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Leu-Asp-Phe-OH
      Purity>95 by HPLC
      SolubilityH₂O (5 mg/ml)
      CommentsMaterials Needed:
      • Protein tyrosine Kinase (such as p60c-src, Cat# PK03)
      • [γ-32P]ATP (10 mCi/ml)
      • Kinase Assay Buffer: 50 mM HEPES, pH 7.5 containing 0.1 mM EDTA and 0.015% Brij® 35 detergent
      • Kinase Dilution Buffer: Kinase assay buffer containing 0.1 mg/mL BSA and 0.2% β-mercaptoethanol (note: b-mercaptoethanol is required for src kinase activity)
      • ATP Mix: 0.15 mM ATP, 30 mM MgCl2 and 200 µCi [γ-32P]ATP per ml in kinase assay buffer
      • Phosphoric acid
      • Acetone
      • P81 Ion Exchange Chromatography paper (i.e. Whatman #3698915)

      Procedure:
      • Dilute kinase protein solution in Kinase Dilution Buffer if of sufficiently high concentration or, alternatively, buffer exchange the kinase protein of interest into the Kinase Dilution Buffer.
      • Mix 10 µl diluted kinase with 10 µl Raytide™ EL (1 mg/ml solution in Kinase Assay Buffer).
      • Start the reaction by adding 10 µl of ATP Mix.
      • Incubate at 30°C for 30 min.
      • Stop the reaction by adding 120 µl of 10% phosphoric acid. Vortex and apply 120 µl onto a square (2 cm x 2 cm) of P81 paper.
      • Wash papers extensively in 0.5% phosphoric acid. Wash once with acetone. Dry and quantitate by liquid scintillation counting. (Phosphorylated Raytide™ EL will stick, while ATP will wash off the papers).
      Storage Avoid freeze/thaw
      +2°C to +8°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C) for long-term storage or refrigerate (4°C) for short-term storage with 0.1% azide. Avoid freeze/thaw cycles of solutions.
      Toxicity Standard Handling