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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

LCAT Activity Assay Kit, Fluorometric Certificates of Analysis

TitleLot Number
428900
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.