Extracting total protein from plant leaf and used a protocol where the lysis buffer is 10%TCA in acetone w/ 0.007g DTT added. The pellet was then re-suspended in acetone and DTT. This also was the wash step. It was a good size pellet when done. However, now I'm trying to dissolve the pellet in water to quantify the proteins using a BCA or bradford assay and I'm having trouble getting the pellet to disolve in the water. Any suggestions.
Well it can be quite difficult, be sure to evaporate acetone as much as you can, after that, you can add your buffer (or water) and you can let it sit at room temperature for like 1h, vortexing it often. You can try to pipet up and down during this time. For difficult samples, I remember using a sonicating bath.
Dissolving the TCA pallet can be tricky. Why water? - Use buffer in which your protein is happy. The last option is you can try like 8M Urea and once dissolved then you can remove urea by dialysis.
If your protein pellet is in acetone, try to air/vacuum dry the pellet to remove all traces of organic solvents. Secondly, since acetone was used, considerable amount of lipids would have come into pellet. Try some basic pH buffer (if desired protein is happy with basic pH) or lower strength NaOH solution to reduce lipids and then resuspend in preferably buffer as Mahesh suggested
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