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Wählen Sie konfigurierbare Panels & Premixed-Kits - ODER - Kits für die zelluläre Signaltransduktion & MAPmates™
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Konfigurierbare Panels & Premixed-Kits
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Kits für die zelluläre Signaltransduktion & MAPmates™
Wählen Sie gebrauchsfertige Kits zur Erforschung gesamter Signalwege oder Prozesse. Oder konfigurieren Sie Ihre eigenen Kits mit Singleplex MAPmates™.
Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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Gewähltes Kit
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96-Well Plate
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Weitere Reagenzien hinzufügen (MAPmates erfordern die Verwendung eines Puffer- und Detektionskits)
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
Platzsparende Option Kunden, die mehrere Kits kaufen, können ihre Multiplex-Assaykomponenten in Kunststoffbeuteln anstelle von Packungen erhalten, um eine kompaktere Lagerung zu ermöglichen.
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Raytide™ EL Substrate: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
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 Number
PK04
Brand Family
Calbiochem®
References
Product Information
Form
Lyophilized
Formulation
Lyophilized from a volatile buffer, BSA.
Applications
Datenkonfigurationsfehler.
Pagelet <collection_feature_application_id-KINASE> nicht gefunden.
Application Comments
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: 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).
Following 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.
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.
Revision
20-February-2008 JSW
Description
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).
Background
Oncogene 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.
Form
Lyophilized
Formulation
Lyophilized from a volatile buffer, BSA.
Recommended reaction conditions
Protocol for Using Raytide in a Kinase AssayMaterials 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).
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: 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 Instructions
Following 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.
