Total Lipid Hydroperoxide ELISA

$725.00

The Total Lipid Hydroperoxide ELISA Assay Kit is intended for the quantitative determination of peroxides in EDTA-plasma, serum and cell culture supernatants. The Total Lipid Hydroperoxide ELISA Assay kit is for research use only and not to be used in diagnostic procedures. 

SKU: PER31-K01 Categories: , ,

Total Lipid Hydroperoxide ELISA

The Total Lipid Hydroperoxide ELISA is For Research Use Only

Size: 1×96 wells
Sensitivity: 6 µmol/L
Incubation Time: 15 minutes
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 10 µl
Controls Included

Product Developed and Manufactured in Germany
Product Support in the United States

Reference Values (Normal)
EDTA-plasma:

< 200 µmol/l    low oxidative stress
200 – 350 µmol/l    moderate oxidative stress
> 350 µmol/l    high oxidative stress

Serum:

< 180 µmol/l    low oxidative stress
180 – 310 µmol/l    moderate oxidative stress
> 310 µmol/l    high oxidative stress

It is suggested that each laboratory establish its own normal ranges


Assay Background

In a healthy body oxidative and reductive processes are in a balance. Free radicals (reactive oxygen species) are eliminated by antioxidants. In case of a lack of antioxidants, free radicals react with cell structures. A reaction with unsaturated fatty acid leads to lipid peroxidation products.  These circumstances are under discussion during the genesis of atherosclerosis. Recent findings suggest that free radicals are also involved in inflammation, sepsis, carcinogenesis and neurodegenerative diseases.
An increased level of peroxides in blood indicates oxidative stress in the body. The measurement of peroxides can indicate an increased risk for heart attack and stroke. A subsequent change in nutrition and/or supplementation therapy might reduce the risk.  The Total Lipid Hydroperoxide ELISA Assay kit allows an easy, rapid and precise quantitative determination of the total peroxides in biological samples. The kit includes all reagents ready to use for preparation of the samples.


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Additional Information

Intra-Assay CV:     2.0 % (168 µmol/l)    [n = 6]
3.6 % (596 µmol/l)    [n = 6]

Inter-Assay CV:    3.6 % (168 µmol/l)    [n = 6]

3.5 % (608 µmol/l)    [n = 6]

Linearity                up to 1000 µmol/l
Detection limit       6 µmol/l

Assay Principle


The determination of the peroxides is performed by the reaction of a peroxidase with peroxides in the sample followed by the conversion of TMB to a colored product.  After addition of a stop solution the samples are measured at 450 nm in a microtiter plate reader. The quantification is performed by the delivered calibrator.

Assay Procedure


  1. Pipette 10 µl Sample, CAL and CTRL in duplicates in appropriate wells.
  2. Pipette 100 µl REABA in appropriate wells.
  3. Measurement 1: Read the absorption of the samples in the ELISA reader at 450 nm.
  4. Add 100 µl reaction buffer mixture to appropriate wells.
  5. Incubate for 15 min at 37° C.
  6. Add 50 µl STOP to appropriate wells.
  7. Measurement 2: Read the absorption immediately after addition of the stop solution (STOP) at 450 nm in the ELISA reader.

Calculation
The difference between measurement 1 and 2 is directly proportional to the peroxide concentration of the sample:
For evaluation, the optical densities of Measurement 1 are subtracted from the optical densities of Measurement 2.
Samples and controls are then calibrated by the use of the calibrator (concentration is given on the label). The concentration of the calibrator is given in H2O2-equivalents (µmol/l).

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