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ÕâÊÇÎÒÒ»Ö±ÓõÄTCA·½·¨
Normal TCA
To eliminate TCA soluble interferences and protein concentration
1) To a sample of protein solution add Trichloroacetic acid (TCA) 100% to get 13% final concentration. Mix and keep 5min ¨C20C and then 15min 4C; or longer time at 4C without the ¨C20C step for lower protein concentration. Suggestion: leave ON if the protein concentration is very low.
1) (preparation of 100% TCA: 454ml H2O/kg TCA. Maintain in dark bottle at 4C.Be careful, use gloves!!!).
2) Spin 15min 4C in microfuge at maximum speed (15000g). Carefully discharge supernatant and retain the pellet: dry tube by inversion on tissue paper (pellet may be difficult to see).
3) For PAGE-SDS, resuspend samples in a minimal volume of sample buffer. (The presence of some TCA can give a yellow color as a consequence of the acidification of the sample buffer ; titrate with 1N NaOH or 1M TrisHCl pH8.5 to obtain the normal blue sample buffer color.) |
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