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428900 LCAT Activity Assay Kit, Fluorometric

LCAT Activity Assay Kit, Fluorometric MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Lecithin:Cholesterol Acyltransferase Activity Assay Kit, Fluorometric

428900
  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityDetection Methods
      HFluorometric
      Description
      Overview

      This product has been discontinued.



      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.
      Catalogue Number428900
      Brand Family Calbiochem®
      SynonymsLecithin:Cholesterol Acyltransferase Activity Assay Kit, Fluorometric
      Application Data
      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.
      Materials Required but Not Delivered•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)
      References
      Product Information
      Detection methodFluorometric
      Form100 Tests
      Kit containsLCAT Substrate, READ Reagent, and a user protocol.
      Quality LevelMQ100
      Applications
      Biological Information
      Assay time4.5 - 8.5 h
      Sample TypePlasma
      Species Reactivity
      • Human
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      R PhraseR: 36/38

      Irritating to eyes and skin.
      S PhraseS: 26

      In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
      Product Usage Statements
      Intended useThe 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.
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Irritant
      Storage Multiple storage requirements
      Storage ConditionsUpon arrival store the LCAT Substrate at -20°C and the READ Reagent at 4°C.
      Protect from Light Protect from light
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Kit containsLCAT Substrate, READ Reagent, and a user protocol.
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      428900 0

      Documentation

      Required Licenses

      Title
      PRODUCTO REGULADO POR LA SECRETARÍA DE SALUD

      LCAT Activity Assay Kit, Fluorometric Certificates of Analysis

      TitleLot Number
      428900

      Brochure

      Title
      Kit SourceBook - 2nd Edition EURO
      Kits SourceBook - 2nd Edition GBP
      User Protocol

      Revision13-July-2012 JSW
      SynonymsLecithin:Cholesterol Acyltransferase Activity Assay Kit, Fluorometric
      Form100 Tests
      Detection methodFluorometric
      Specieshuman
      StorageUpon arrival store the LCAT Substrate at -20°C and the READ Reagent at 4°C.
      Intended useThe 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.
      Principles of the assayLCAT 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.
      Materials providedThere 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
      Materials Required but not provided•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)
      Reagent preparation•READ Reagent: reconstitute the vial of READ Reagent with 29.7 ml READ Reagent Reconstitution Buffer.
      Detailed protocol1. 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.
      Example data

      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.
      Registered TrademarksCalbiochem® is a registered trademark of Merck KGaA, Darmstadt.
      Interactive Pathways™ is a trademark of Merck KGaA, Darmstadt.