Vitamin E (VE) Colorimetric Assay Kit

Catalogue Number: E-BC-K033-S-ELA

Manufacturer:	Elabscience
Shelf Life:	6 months
Type:	Detection Kit
Shipping Condition:	Blue Ice
Storage Condition:	2-8°C
Unit(s):	100 assays, 50 assays
Range:	0.09-40 µg/mL
Sensitivity:	0.09 µg/mL
Sample type:	Plasma, Serum, Tissue
Sample size:

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Description

Description: Application This kit can be used for detection of VE content in samples, such as animal/plant tissue, serum (plasma). Detection significance VE is a kind of natural lipid-soluble antioxidant, which exist in cellular membrane structure (cell membrane, mitochondrial, microsomal membrane), lipid droplets of adipocytes and lipoproteins of plasma. It is scavenger of singlet oxygen and superoxide, blocking agent of lipid peroxidation and can protect protein sulfhydryl. Detection principle Fe3+ can be deoxidized to Fe2+ by VE with ferroin existing. Fe2+ can react with phenanthroline and form pink compound under certain condition. After colorimetric assay, VE content can be figured out according to the standard curve or calculated through formula. Experimental instrument Test tube, Micropipettor, Vortex mixer, Centrifuge, Spectrophotometer (533 nm) Operation steps It is recommended to take 2~3 samples which expected large difference to do pre-experiment before formal experiment. 1. Detection of VE content in serum (plasma) (1) Extraction of n-heptane VE in serum (plasma) (n-heptane is self-prepared): Vortex mixing (extract thoroughly) for 1 min, then centrifuge at 3500~4000 rpm for 5~10 min. Take the n-heptane VE extraction solution (the upper layer solution) for chromogenic reaction. Note: Liquid in the tube will be divided into three layers, and the upper layer is n-heptane VE extraction solution, the middle layer is water and absolute ethyl alcohol and the lower layer is protein precipitate. (2) Chromogenic reaction: Mix fully and stand for 2 min. Measure the OD values of each tube at 533 nm wavelength with cuvette of 0.5 cm optical path, set to zero with absolute ethyl alcohol. 2. Detection of VE content in tissue (1) Sample pretreatment: Preparation of 10% homogenate solution: weigh the tissue accurately. Add Reagent 4 (homogenate medium) in a weight (g): volume (mL) ratio of 1: 9, then use mechanical homogenization in ice water bath. Centrifuge the mixture solution at 2500 rpm for10 min and take the supernatant for measurement. (2) Extraction of n-heptane VE in tissue homogenate (n-heptane is not provided in this kit): 1.0

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2-8°C

E-BC-K033-S Protocol

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£128.00

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