Yesterday i faced to a problem while diluting my sample, Let me give you a background of what i did,
My plant protein (seed) was extracted through TCA-acetone methanol washes and phenol precipitation (The protocol has been attached)
19 downloads, then solubilised in dense SDS buffer (30% sucrose, 2% SDS,0.1 M Tris-HCl, pH 8.0, 5% 2-mercaptoethanol). I had a considerable amount of pellet, so i added 500Ml (Microliter) of SDS buffer. The whole pellet was easily dissolved. then i run SDS-PAGE. Before performing SDS-PAGE i prepared dilution 10-20-30 of my protein.while adding water i saw no turbidity and everything was normal, no turbidity, no precipitation.....but in the second experience i solubilised another pellet, but i solubilised it with 400ML, it solubilised easily again But when i started to prepare dilutions, while adding water, i saw considerable turbidity and after minutes the protein precipitated I couldn't make dilutions. Why did this happen? Why this happend to second experience while i had no problem with dilution in the first experience. If there is any blur, please let me know to elaborate.
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