Wheat storage proteins can be obtained with alcoholic solutions (70% ethanol or 50% propanol-1) containing 2-mercaptoethanol or dithiothreitol. Following, you should precipitate this solution with cold-acetone and to resuspend in a buffer solution as 625 mM Tris-HCl pH: 6.8.

Thanks Dear Alvarez for your opinion.

But by using this Tris-HCl solution, Gluten protein of wheat are not solubilise and even it was not solubilised in 1X-PBS, 1% DMSO, and 0.1 M Acetic acid also.

So, please tell me some other solvent.

Thanks

Gluten proteins (glutenins and gliadins) are solubilized in this buffer.

Gliadins, the main responsible of celiac disease, are soluble in 70% ethanol.

The natural polymers of glutenins are not soluble in this solution, but if you added 2% dithiothreitol in the extraction solution (step 1), these polymers are denatured and their subunits are soluble in Tris-HCl buffer.

we can not use 70% Ethanol in cell lines, this will denature the cell proteins and result into adverse effects on caco2 cells.

Dear Kunj,

I may be late to answer but gliadins are soluble in DMSO. Need not to use 1% DMSO, instead use Pure DMSO or 50% of it.

So that 50mg gliadins can be dissolved in 1ml DMSO that can be further diluted below it's toxic limits for cell culture.