428900 | LCAT Activity Assay Kit, Fluorometric

428900
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      Overview

      Key Spec Table

      Nachweismethoden
      Fluorometric

      Preis & Verfügbarkeit

      BestellnummerVerfügbarkeit Verpackung St./Pkg. Preis Menge
      428900-1KIT
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          Beschreibung
          Übersicht Assay kit useful for the quantitative measurement of lecithin:cholesterol acyltransferase activity (LCAT) in human plasma. LCAT mediates the formation of cholesteryl esters in human plasma by transferring an acyl chain from the sn-2 position of phosphatidylcholine to cholesterol.
          Bestellnummer 428900
          Produktreihe Calbiochem®
          Synonyme Lecithin:Cholesterol Acyltransferase Activity Assay Kit, Fluorometric
          Anwendungsdaten
          The non-hydrolyzed LCAT Substrate fluoresces at 470 nm. Upon hydrolysis by LCAT, a monomer is released from the LCAT Substrate that fluoresces at 390 nm. LCAT activity is assessed as a change in 470/390 emission intensity.

          Normal human plasma is assayed according to the Detailed Protocol above. Over time the 470/390 ration decreases, indicating LCAT activity in the sample.
          Literatur
          Produktinformationen
          Nachweismethode Fluorometric
          Form 100 Tests
          Kitinhalt LCAT Substrate, READ Reagent, and a user protocol.
          Anwendungen
          Biologische Informationen
          Testzeit 4.5 - 8.5 h
          Probenart Plasma
          Spezies human
          Physiochemische Informationen
          Abmessungen
          Materialinformationen
          Angaben zur Toxikologie
          Sicherheitsinformationen gemäß GHS
          Sicherheitshinweise
          R-Satz R: 36/38

          S-Satz S: 26

          Hinweise zum Produktgebrauch
          Verwendungszweck The Calbiochem® LCAT Activity Assay Kit, Fluorometric is intended for the quantitative measurement of lecithin:cholesterol acyltransferase activity (LCAT) in normal human plasma. This kit allows the quantitation of fold of increase or decrease of enzymatic activity among samples, this kit also allows the quantitation of the % change of 470/390 ratio with time, which can be used to define LCAT enzymatic activity. This kit does not allow the measurement of substrate concentration or the amount of hydrolyzed substrate.
          Lagerungs- und Versandinformationen
          Versandcode Blue Ice Only
          Toxizität Irritant
          Lagerung Multiple storage requirements
          Lagerungsbedingungen Upon arrival store the LCAT Substrate at -20°C and the READ Reagent at 4°C.
          Vor Licht schützen Protect from light
          Nicht einfrieren No
          Verpackungsinformationen
          Transportinformationen
          Ergänzende Informationen
          Kitinhalt LCAT Substrate, READ Reagent, and a user protocol.
          Spezifikationen

          Documentation

          SDB

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          Analysenzertifikate

          TitelChargennummer
          428900

          Broschüre

          Titel
          Kit SourceBook - 2nd Edition EURO
          Anwenderprotokoll

          Revision 13-July-2012 JSW
          Synonyme Lecithin:Cholesterol Acyltransferase Activity Assay Kit, Fluorometric
          Form 100 Tests
          Nachweismethode Fluorometric
          Spezies human
          Lagerung Upon arrival store the LCAT Substrate at -20°C and the READ Reagent at 4°C.
          Verwendungszweck The Calbiochem® LCAT Activity Assay Kit, Fluorometric is intended for the quantitative measurement of lecithin:cholesterol acyltransferase activity (LCAT) in normal human plasma. This kit allows the quantitation of fold of increase or decrease of enzymatic activity among samples, this kit also allows the quantitation of the % change of 470/390 ratio with time, which can be used to define LCAT enzymatic activity. This kit does not allow the measurement of substrate concentration or the amount of hydrolyzed substrate.
          Grundlagen des Tests LCAT mediates the formation of cholesteryl esters in human plasma by transferring an acyl chain from the sn-2 position of phosphatidylcholine to cholesterol. Using the LCAT Activity Assay, Fluorometric, human plasma samples are incubated with LCAT Substrate, a fluorescent substrate that emits fluorescence at 470 nm. When the substrate is hydrolyzed by LCAT, a monomer is released that emits fluorescence at 390 nm. The LCAT activity is assessed as a change in 470/390 emission intensity.










          Figure 1: Assay Principle

          The non-hydrolyzed LCAT Substrate fluoresces at 470 nm. Upon hydrolysis by LCAT, a monomer is released from the LCAT Substrate that fluoresces at 390 nm. LCAT activity is assessed as a change in 470/390 emission intensity.

          Mitgelieferte Materialien There are sufficient reagents for 100 tests.

          •LCAT Substrate (Kit Component No. KP23001): 1 vial, 100 µl
          •READ Reagent (Kit Component No. KP23002): 1 vial, 350 µl
          Erforderliche, jedoch nicht mitgelieferte Materialien •LCAT Assay Buffer (150 mM NaCl, 10 mM Tris-HCl, 4 mM 2-mercaptoethanol, 1 mM EDTA, 7.4)
          •READ Reagent Reconstitution Buffer (150 mM NaCl, 10 mM Tris-HCl, 1 mM EDTA, pH 7.4)
          Ansetzen von Reagenzien •READ Reagent: reconstitute the vial of READ Reagent with 29.7 ml READ Reagent Reconstitution Buffer.
          Ausführliches Protokoll 1. Mix 1 µl LCAT Substrate 200 µl LCAT Assay Buffer with sample containing LCAT (e.g., 3-5 µl normal human plasma) and incubate for 4-8 h at 37°C.
          2. Add 100 µl of the mixture in step 1 to 300 µl READ Reagent and vortex. Read the fluorescence at an excitation wavelength of 340 nm and an emission wavelength of 390 nm and 470 nm. DO NOT incubate the samples in the presence of READ Reagent, as this will inactivate the LCAT in the sample.

          NOTE: The emission intensities at 390 nm and 470 nm represent the emission of the substrate hydrolyzed (390 nm) and not hydrolyzed (470 nm). In the presence of LCAT, a ratio of the two emission intensities (470/390) will indicate an increase in concentration of the monomer, which emits at 390 nm, and the simultaneous decrease in concentration of the non-hydrolyzed LCAT Substrate, which emits at 470 nm.
          Beispieldaten

          Figure 2: Example Data

          Normal human plasma is assayed according to the Detailed Protocol above. Over time the 470/390 ration decreases, indicating LCAT activity in the sample.

          Eingetragene Marken Calbiochem® is a registered trademark of Merck KGaA, Darmstadt.
          Interactive Pathways™ is a trademark of Merck KGaA, Darmstadt.