Questions on TBARS assay for lipid peroxidation

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hanhan2008
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Questions on TBARS assay for lipid peroxidation

Post by hanhan2008 » Sat Dec 01, 2007 11:03 pm

HI,

I've been trying the method using either cell culture or blood plasma.

However,
1. The cell lysate, after reaction with TBA, gave a yellow color, while the tetraethoxypropane standards were in pink. Absorbance spectrum showed much bigger peaks in other locations than the peak at desired location of 532 nm.
2. I tried H2O2 as a positive control and didn't get increase over control.
3. Based on my online literature research, there seems too many variations to this method: (w/ or w/o SDS, butanol extraction, etc. For sample homogenization alone, there can be sonication, glass homogenizor, or using tris-HCl (pH=7.4)

Could anybody suggest which is the best way to do it (before I have to turn to HPLC method)? I'm under increasing pressure of making progress, so any help will be highly appreciated.

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