TCA precipitates macromolecules.

Assay of TBARS (thiobarbituric acid reactive substances) measures concentration of malondialdehyde produced due to degradation of unstable lipid peroxides.

TCA has two functions first dehydrating the hydration shells around the protein by which the anionic TCA triggers partial protein unfolding through disruption of the electrostatic interactions by determining the native tertiary structure of the protein. TCA is less effective for disordered proteins.

hii Mateus,

TCA is an extracting agent. It is preferred for its ability to precipitate the protein.

TCA precipitates macromolecules.

Assay of TBARS (thiobarbituric acid reactive substances) measures concentration of malondialdehyde produced due to degradation of unstable lipid peroxides.

TCA has two functions first dehydrating the hydration shells around the protein by which the anionic TCA triggers partial protein unfolding through disruption of the electrostatic interactions by determining the native tertiary structure of the protein. TCA is less effective for disordered proteins.

TCA added for TBARS assayis in purpose to accelerate condensation reaction of reagent TBA with MDA, since TCA is stronger acid compare with acetic acid, thus more color of chromophore (red) was performed well.

I do agree with the above statements regarding the action of TCA in the assay mixture of TBARS

Dear all,

I'm struggling a lot with this method! I'm trying to implement TBARS using Caco-2 cell line (allowed to growth for 7 days, next a 24 hour incubation with my samples of study). My real problem is how to lysate cells. I've tried 10% and 20% (w/v --> 500 uL) TCA but I don't get an homogenize solution (after incubationg 30 minutes on ice). Should be like this or should look and homogenize solution.

The following procedure is to add 500 uL TBA (0.1%) and heat 10 minutes at 80-90ºC.

Alternatively, I've try higher concentrations of TCA (until 70% w/v) which seems easily to homogeneize, however, didn't get an quantification for bradford... Please help me! Btw I don't have sonicator or ultraturrax, thus, must find easy way to lysate cells.

Thanks in advance