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PK04 Raytide™ EL Substrate

Raytide™ EL Substrate MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
PK04
  
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Overview

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
Catalogue Number GTIN
PK04 0

Documentation

Raytide™ EL Substrate Certificates of Analysis

TitleLot Number
PK04

Brochure

Title
Protein Phosphatases Technical Bulletin
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.

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