b******e
发帖数: 3348 | 1 我只做过cell culture,对mouse model一窍不通，
做肿瘤westernblot的话是不是可以可以和细胞一样直接在sds-dtt buffer里
homogenize sample,煮沸 然后run, | w*****3
发帖数: 1582 | 2 我做过人的
For preparation of protein extracts, renal tumor tissue was pulverized with
a mortar under liquid nitrogen and suspended on ice in lysis buffer (20 mM
Hepes, pH 7.7, 0.2 M NaCl, 1.5 mM MgCl2, 0.4 mM EDTA, 1% Triton X-100, 0.5
mM DTT, 100 µg/ml leupeptin, 100 µg/ml aprotinin, 10 mM
benzamidine, 2 mM phenylmethylsulphonyl fluoride, 20 mM -
glycerophosphate, and 0.1 mM sodium-orthovanadate).
然后加 sds-dtt buffer里 homogenize sample,煮沸 然后run,
跟细胞不同的是非常容易degradation. | b******e
发帖数: 3348 | 3 非常感谢！how about RNA extraction
with
【在 w*****3 的大作中提到】
: 我做过人的
: For preparation of protein extracts, renal tumor tissue was pulverized with
: a mortar under liquid nitrogen and suspended on ice in lysis buffer (20 mM
: Hepes, pH 7.7, 0.2 M NaCl, 1.5 mM MgCl2, 0.4 mM EDTA, 1% Triton X-100, 0.5
: mM DTT, 100 µg/ml leupeptin, 100 µg/ml aprotinin, 10 mM
: benzamidine, 2 mM phenylmethylsulphonyl fluoride, 20 mM -
: glycerophosphate, and 0.1 mM sodium-orthovanadate).
: 然后加 sds-dtt buffer里 homogenize sample,煮沸 然后run,
: 跟细胞不同的是非常容易degradation.
| G****a
发帖数: 280 | 4 you could also try use higer concentration Urea solution, you even do use to
boiling before load to SDS-PAGE |
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