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devilpanda

金虫 (小有名气)

应助: 0 (幼儿园)
金币: 386.5
红花: 1
帖子: 105
在线: 37.5小时
虫号: 1410385
注册: 2011-09-21
性别: GG
专业: 天然药物化学

[求助] 小弟求助

Whole-cell extracts were prepared by
lysing the cells with RIPA buffer containing 150 mM NaCl, 50 mM
Tris HCl (pH 8), 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS,
and protease inhibitor and phosphatase inhibitor cocktails (Sigma-
Aldrich). The cell extracts were separated by 10% SDS-PAGE and
transferred onto a polyvinylidene fluoride membrane (Millipore,
Billerica, MA, USA). Samples were incubated in blocking buffer (0.1%
Tween 20 and 5% nonfat milk powder in Tris-buffered saline [TBS])
for 1 h at room temperature. Afterward, the membrane was incubated
with primary antibody in blocking buffer overnight at 4 °C before
being washed twice with TBST (0.1% Tween in TBS) and incubated
with the appropriate secondary antibody in blocking buffer for 1 h at
room temperature. The blot was developed using ECL Western
blotting substrate (Millipore) and analyzed using a luminescent image
analyzer (LAS-4000 mini; FujiFilm, Tokyo, Japan). The primary
antibodies were used at the following dilutions: rat anti-ABCG2, 1:100
(Abcam, Cambridge, UK); rabbit anti-PARP, 1:1000 (Cell Signaling
Technology, Beverly, MA, USA); rabbit anti-caspase 7, 1:1000 (Cell
Signaling Technology); rabbit anti-Akt and phosphate Akt-ser 473,
1:1000 (Cell Signaling Technology); and mouse anti-β-actin, 1:10000
(Sigma-Aldrich)谁能帮我翻译下啊~~