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chunqizzm

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[ Last edited by chunqizzm on 2012-3-25 at 23:36 ]
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chunqizzm: ½ð±Ò+5 2012-03-27 07:21:19
chunqizzm: »ØÌûÖö¥ 2012-03-27 07:21:45
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6Â¥: Originally posted by zxc6884 at 2012-03-26 21:07:16:
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 concentr ...

Õâ¸öÊÇ×î³£ÓõÄTCA³Áµí·½·¨£¬Ò»°ãÐèÒªµ°°×ÔÚ5¦Ìg/mlÒÔÉϲÅÄܳÁÏÂÀ´£¬Ò²¾ÍÊÇ˵´ó¸Å»¹ÓÐ5¦Ìg/ml×óÓÒµ°°×²»ÄܳÁµíÀ´£¬ÕâÊÇÀíÂ۵ģ¬Êµ¼ÊÊÕÂÊÓëÑùÆ·ºÍ²Ù×÷Ïà¹Ø£¬Èç¹ûÄãÑùÆ·±ÈÕâŨ¶È»¹µÍµÄ»°¿ÉÒÔÊÔÏÂTCA-DOC³Áµí£¬ÄܳÁµí1¦Ìg/mlÒÔϵĵ°°×£¬È»ºóÒøÈ¾¡£

TCA-DOC
For precipitation of very low protein concentration
1) To one volume of protein solution, add 1/100 vol. of 2% DOC (Na deoxycholate, detergent).
2) Vortex and let sit for 30min at 4C.
3) Add 1/10 of Trichloroacetic acid (TCA) 100% vortex and let sit ON at 4C (preparation of 100% TCA: 454ml H2O/kg TCA. Maintain in dark bottle at 4C.Be careful, use gloves!!!).
4) 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). [OPTION: Wash pellet twice with one volume of cold acetone (acetone keep at ¨C20C). Vortex and re-pellet samples 5min at full speed between washes].
5) Dry samples under vacuum (speed vac) or dry air. For PAGE-SDS, resuspend samples in a minimal volume of sample buffer. (The presence of some TCA can give a yellow colour 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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9Â¥2012-03-26 22:04:27
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chunqizzm

Òø³æ (СÓÐÃûÆø)


ÒýÓûØÌû:
2Â¥: Originally posted by zlu520 at 2012-03-25 23:17:06:
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3Â¥2012-03-25 23:43:09
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mingxie

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4Â¥2012-03-26 09:13:32
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xingnian0925

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