TCA-丙酮蛋白浓缩
TCA protein precipitation protocol
Stock Solutions: 100% (w/v) Trichloroacetic acid (TCA)
recipe: dissolve 500g TCA (as shipped) into 350 ml dH2O, store at RT.
Precipitation Protocol:
1. Add 1 volume of TCA stock to 4 volumes of protein sample.
i.e. in 1.5ml tube with maximum vol., add 250µl TCA to 1.0ml sample.
2. Incubate 10 min at 4°C.
3. Spin tube in microcentrifuge at 14K rpm, 5 min.
4. Remove supernatant, leaving protein pellet intact. Pellet should be formed from whitish,fluffy ppt.
5. Wash pellet with 200µl cold acetone.
6. Spin tune in microfuge at 14K rpm, 5min.
7. Repeat steps 4-6 for a total of 2 acetone washes.
8. Dry pellet by placing tube in 95°C heat block for 5-10 min to drive off acetone.
9. For SDS-PAGE, add 2X or 4X sample buffer (with or without bME) and boil smaple for
10 min in 95°C herat block before loading smaple onto polyacrylamide gel.
TCA蛋白浓缩步骤：
储存液：100%（W/V）三氯乙酸（TCA）
配制：将500g TCA溶解到350ml dH2O中，室温储存。

浓缩步骤：
1.加1倍体积的TCA储存液到4倍体积的蛋白样品中。

如：在1.5ml的离心管中，1ml
样品中加入250ul的TCA。

2.4度孵育10min。

3.14 000rpm 离心5 min。

4.弃上清
5.加入200ul预冷的丙酮洗涤沉淀。

6.14 000rpm 离心5 min。

7.重复步骤4-6两次。

8.95度5-10 min，晾干沉淀以彻底出去丙酮。

9.加入2X或4X样品buffer，煮沸样品10 min，SDS-PAGE。