Question

(5)求论文翻译,急!!!

2.7. Western blotting analysis
Acid urea–PAGE and Western blotting analyses were conducted in
accordance with the method described by Panyutich and Ganz [24]. In
the Western blotting experiments, we used an antiserum that had previously
been generated via an injection of 18Hc into a rabbit [25]. In
brief, the samples were electrophoresed on acid urea–PAGE gels and
transferred to nitrocellulose membranes in Tris–glycine buffer
(25 mM Tris, 192 mM glycine, 20% methanol, pH 8.3) at 100 V for
1 h. After transference, the membranes were equilibrated in Tris–buffered
saline (20 mM Tris, 500 mM NaCl, pH 7.5; TBS) for 10 min
and then incubated for 30 min in blocking solution (3% skim milk in
TBS). After a brief washing in TBS containing 0.05% Tween-20
(TTBS), the membrane was incubated for 1 h in a primary antibody
solution (antiserum diluted 100-fold with blocking solution). The
membrane was washed twice in citrate-buffered saline (20 mM citrate,
500 mM NaCl, pH 5.5) containing 0.05% Tween-20, for 5 min per
washing, and then incubated for 1 h in blocking solution containing
a 3000-fold diluted secondary antibody (GAR-HRP conjugated IgG;
Bio-Rad, Richmond, CA). Finally, the membrane was washed twice
in TTBS, once in TBS, and finally submerged in HRP color development
solution (50 ml of TBS containing 30 ll of H2O2 mixed with
0.3% of chloronaphthol in 10 ml of ice-cold methanol).

Answer 1

2.7 。蛋白质印迹分析
酸尿素- PAGE和免疫印迹分析进行了在
按照该方法所描述的panyutich和冈茨[ 24 ] 。在…内
该免疫印迹实验中，我们使用了抗说，以前曾
产生了通过注资18hc成兔[ 25 ] 。在…内
简单来说，样本electrophoresed对酸性尿素页凝胶和
转移到硝化纤维素膜在三-甘氨酸缓冲
（ 25毫米三，一百九十二毫米甘氨酸， 20 ％的甲醇， pH值8.3 ）在100伏为
1 h后划转后，膜平衡在三缓冲
生理盐水（ 20毫米三， 500毫米氯化钠， pH值7.5 ;汤匙）为10分钟
然后孵育30分钟，在阻塞的解决办法（ 3 ％脱脂牛奶在
东京广播公司） 。经过短暂洗衣机在汤匙含有0.05 ％ ，吐温-20
（ ttbs ） ，膜孵育1 h后在一小学抗体
解决方案（抗血清稀释100倍，与拦截的解决方案） 。那个
膜被洗净，两次在柠檬酸缓冲液（ 20毫米的柠檬酸，
500毫米氯化钠， pH值5.5 ） ，其中载有0.05 ％ ，吐温-20 ，为5分钟％
洗衣机，然后孵育1 h后，在封锁的解决方案包含
1 3000倍稀释中学抗体（明嘉-辣根过氧化物酶共轭IgG抗体;
生物的RAD ，里士满，加州） 。最后，膜的清洗两次
在ttbs ，一旦在东京广播公司，最后淹没在HRP的彩色发展
解决方案（ 50毫升的汤匙载有30人当地雇员的过氧化氢混合
0.3 ％ chloronaphthol在10毫升的冰冷的甲醇） 。

Answer 2

不会 啊啊~