Human TNF alpha (Colorimetric)

[ Home ] [ Up ] [ Home ] [ Up ] HDL/LDL Electrophoresis Albumin—OsrHSA Pricelist: http://www.diagrade.com/search.php

Free Live Chat Software

OUR SUPPLIERS

KOMABIOTECH
301, Gayang Technotown, #1487 Gayang 3 dong, Gangseo-gu
Seoul 157-793, KOREA

Spherotech, Inc.
27845 Irma Lee Circle, Unit 101
Lake Forest, IL 60045

Exalpha Biologicals, Inc.
2 Shaker Road, Unit B101
Shirley, MA 01464

SCETI K.K
BIOSCIENCE Export DF Kasumigaseki Place,3-6-7 Kasumigaseki Chiyoda-ku, Tokyo 100-0013 JAPAN

EY Laboratories, Inc. Headquarters
107 N. Amphlett Blvd
San Mateo, CA. 94401 USA

EXBIO Praha, a.s.
Nad Safinou II 366
252 42 Vestec
Czech Republic

Sacace Biotechnologies S.r.l.
Via Scalabrini, 44
22100 Como Italy

GENTAUR BVBA

VAT BE0473327336

Av. de l Armee 68 B4

1040 Brussels BELGIUM

Tel + 32 16 58 90 45

Fax + 32 16 50 90 45

GENTAUR France SARL

SIRET 48423788800017

Rue Lagrange, 9

75005 Paris, France

Tel 01 43 25 01 50

Fax 01 43 25 01 60

GENTAUR Germany Marienbongard 20

52074 Aachen, Germany

Tel 0241 56 00 99 68 Fax 0241 56 00 47 88

GENTAUR Pol Sp. Z.o.o. Ulica Ogarna 15/19B m2

80-826 GDANSK

Tel 00 48 58 760 77 08

Fax: 00 32 16 50 90 45

GENTAUR Italy

23015 Milano, Italy

Tel 02 36 00 65 93

Fax 02 36 00 65 94

Česká republika Praha
+420246019719

Danmark

+4569918806

Finland Helsset
+358942419041

Ελλάς Αθήνα
+302111768494

Ireland Dublin
+35316526556

Luxembourg
+35220880274

Magyarország Budapest
+3619980547

Nederland
+31208080893

Norge Oslo
+4721031366

Österreich
+43720880899

Sverige Stockholm
+46852503438

Schweiz Züri
+41435006251

Northern America

Canada Montreal
+15149077481

US New York
+17185132983

Asia 日本
+81 78 386 0860

Other Countries
0032 (0)16 41 44 07

EnzyChromTM AF HDL and LDL/VLDL Assay Kit (E2HL-100)

Quantitative Colorimetric/Fluorimetric Determination of HDL and LDL/VLDL

DESCRIPTION

CHOLESTEROL concentrations in High-Density Lipoprotein (HDL) and Low-Density (LDL)/Very-Low-Density (VLDL) Lipoproteins are strong predictors for coronary heart disease. Functional HDL offers protection by removing cholesterol from cells and atheroma. Higher concentrations of LDL and lower concentrations of functional HDL are strongly associated with cardiovascular disease due to higher risk of atherosclerosis. The balances between high- and low-density lipoproteins are solely genetically determined, but can be changed by medications, food choices and other factors. Simple, direct and automation-ready procedures for measuring HDL and LDL/VLDL concentrations are very desirable. BioAssay Systems' HDL and LDL/VLDL quantification kit is based on our improved PEG precipitation method in which HDL and LDL/VLDL are separated, and cholesterol concentrations are determined using a single Working Reagent that combines cholesterol ester hydrolysis, oxidation and color reaction in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at λem/ex = 585/530nm is directly proportional to total cholesterol concentration in the sample.

APPLICATIONS

Direct Assays: HDL and LDL/VLDL cholesterol in serum samples.

Pharmacology: evaluation of drugs on cholesterol metabolism.

KEY FEATURES

Sensitive and accurate. Linear detection range in 96-well plate: 1 to 100 mg/dL cholesterol for colorimetric assays and 0.2 to 10 mg/dL for fluorimetric assays.

Convenient. Room temperature assay. No 37°C heater is needed.

KIT CONTENTS (100 assays in 96-well plates)

PBS: 2 x 1.5 mL Precipitation Reagent: 1.5 mL

Assay Buffer: 20 mL Enzyme Mix: 120 uL

Dye Reagent: 120 µL Standard: 1 mL 300mg/dL cholesterol

Storage conditions. Store PBS and Precipitation Reagent at room temperature and the rest reagents at -20°C. Shelf life of at least 6 months (see expiry dates on labels).

Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.

COLORMETRIC PROCEDURES

Important: bring all reagents except enzyme mix to room temperature prior to assay. Non-hemolyzed serum samples should be used.

1. Sample Preparation. Transfer 20µL serum into a 1.5-mL centrifuge tube, add 20 µL Precipitation Reagent. Vortex to mix and centrifuge 5 min at 9,500 x g (e.g. 9,500 rpm in an Eppendorf 5415C tabletop centrifuge). Carefully transfer 24 µL supernatant into a clean tube, add 96 µL Assay Buffer. Label this tube “HDL”. Carefully remove all remaining supernatant from the pellet. Transfer 40 µL PBS to the pellet and mix by repeated pipetting. Transfer 24 µL mixture into another clean tube, add 96 µL Assay Buffer. Label this tube “LDL/VLDL”. In a third tube, transfer 12 µL serum sample and mix well with 108 µL Assay Buffer. Label this tube “Total”. Cholesterol Standard: transfer 5 µL 300 mg/dL cholesterol and mix with 145 µL Assay Buffer. Label this tube “Standard”.

2. Assay. Transfer 50 µL Assay Buffer (“Blank”), 50 µL Standard, 50 µL “Total”, 50µL “HDL” and 50 µL “LDL/VLDL” into wells of a clear flat-bottom 96-well plate. If desired, run assays in duplicate. For each reaction well, mix 55 µL Assay Buffer with 1µL Enzyme Mix and 1 µL Dye Reagent. Add 50 µL of this Working Reagent to each standard and sample well. Tap plate to mix well. Incubate 30 min at room temperature. Read OD values at 570 nm.

Note: if the Sample OD is higher than the Standard OD, dilute sample in assay buffer and repeat the assay. Multiply result by the dilution factor.

3. Calculation. Cholesterol concentrations in the Total, HDL and (LDL/VLDL) fractions are calculated as follows,

FLUORIMETRIC PROCEDURE

Dilute the Samples and Standard prepared in Colorimetric Procedure 1:10 in Assay Buffer. Transfer 50 µL diluted standards and 50 µL diluted samples into separate wells of a black 96-well plate. Add 50 µL Working Reagent (see Colorimetric Procedure). Tap plate to mix. Incubate 30 min at room temperature and read fluorescence at λex = 530nm and λem = 585nm.

Note: if the Sample F is higher than the Standard F, dilute sample in assay buffer and repeat the assay. Multiply result by the dilution factor. The cholesterol concentration of Sample is calculated as

MATERIALS REQUIRED, BUT NOT PROVIDED

Pipetting devices, 96-well plate and plate reader.

EXAMPLES

Serum samples were run in duplicate according to the standard procedure.

LITERATURE

[1]. Viikari J. (1976). Precipitation of plasma lipoproteins by PEG 6000 and its evaluation with electrophoresis and ultracentrifugation. Scand J Clin Lab Invest 36:265-268.

[2]. Demacker, PMN et al. (1980). A study of the use of polyethylene glycol in estimating cholesterol in high density lipoprotein. Clin Chem 26:1775-1779.

[3]. Widhaim, K. and Pakosta, R. (1991). Precipitation with Polyethylene Glycol and Density-Gradient Ultracentrifugation Compared for Determining High-Density Lipoprotein Subclasses HDL2 and HDL3. Clin. Chem 37/2, 238-240.

EnzyChromTM HDL and LDL/VLDL Assay Kit (EHDL-100)

Quantitative Colorimetric Determination of HDL and LDL/VLDL Cholesterol

DESCRIPTION

Simple, direct and automation-ready procedures for measuring HDL and LDL/VLDL concentrations are very desirable. BioAssay Systems' HDL and LDL/VLDL quantification kit is based on our improved PEG precipitation method in which HDL and LDL/VLDL are separated, and cholesterol concentrations are determined using cholesterol esterase/cholesterol dehydrogenase reagent. In this reaction, NAD is reduced to NADH. The optical density of the formed NADH at 340 nm is directly proportionate to the cholesterol concentration in the sample.

APPLICATIONS

Direct Assays: HDL and LDL/VLDL cholesterol in serum samples from any species.

Pharmacology: evaluation of drugs on cholesterol metabolism.

KEY FEATURES

Sensitive and accurate. Requires only 20 μL serum sample.

Detection limit of 5 mg/dL, linearity up to 300 mg/dL cholesterol in 96- well plate assay.

Convenient. Room temperature assay. No 37°C heater is needed.

KIT CONTENTS (100 assays in 96-well plates)

PBS: 1.5 mL Precipitation Reagent: 1.5 mL

Assay Buffer: 20 mL Enzyme Mix: 120 uL

NAD Solution: 2 mL Standard: 1 mL 300mg/dL cholesterol

Storage conditions. Store PBS and Precipitation Reagent at room temperature and the rest reagents at -20°C. Shelf life of at least 6 months (see expiry dates on labels).

PROCEDURES

Important: bring all reagents except enzyme mix to room temperature prior to assay. Non-hemolyzed serum samples should be used. The following procedure is designed for duplicate determinations.

1. Sample Preparation. Transfer 20 μL serum into a 1.5-mL centrifuge tube, add 20 μL Precipitation Reagent. Vortex to mix and centrifuge 5 min at 9,500 x g (e.g. 9,500 rpm in an Eppendorf 5415C tabletop centrifuge). Carefully transfer 24 μL supernatant into a clean tube, add 96 μL Assay Buffer. Label this tube “HDL”. Carefully remove all remaining supernatant from the pellet. Transfer 40 μL PBS to the pellet and mix by repeated pipetting. Transfer 24 μL mixture into another clean tube, add 96 μL Assay Buffer. Label this tube “LDL/VLDL”. In a third tube, transfer 12 μL serum sample and mix well with 108 μL Assay Buffer. Label this tube “Total”. Cholesterol Standard: transfer 12 μL 300 mg/dL cholesterol and mix with 108 μL Assay Buffer. Label this tube “Standard”.

2. Assay. Transfer 50 μL Assay Buffer (“Blank”), 50 μL Standard, 50 μL “Total”, 50μL “HDL” and 50 μL “LDL/VLDL” into wells of a clear bottom 96-well plate. If desired, run assays in duplicate. Prepare enough Working Reagent. For each reaction well, mix 50 μL Assay Buffer, 18 μL NAD Solution and 1 μL Enzyme Mix. Transfer 60 μL of the Working Reagent to each reaction well. Tap plate to mix well. Note: addition of Working Reagent to all wells should be rapid and mixing should be thorough. Use of a multichannel pipettor is recommended. Incubate 30 min at room temperature. Read OD values at 340nm.