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xinyuecao123

新虫 (初入文坛)

[求助] ELISA

我用简介ELISA测多克隆抗体效价,显色正常逐级递减,但测的值阴性和阳性还有空白对照却相差不大,且空白对照也显色了,有哪位高人指点一下,这是什么原因,不胜感激
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xl_dut

木虫 (正式写手)

【答案】应助回帖

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gyesang: 金币+2, 鼓励回帖交流! 2013-10-01 17:00:34
可能是加抗体前封闭不彻底或者加抗体后洗脱不充分。空白对照也显色的原因极有可能是封闭不彻底!
一日之计在于昨天!
2楼2013-10-01 13:29:06
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xinyuecao123

新虫 (初入文坛)

引用回帖:
2楼: Originally posted by xl_dut at 2013-10-01 13:29:06
可能是加抗体前封闭不彻底或者加抗体后洗脱不充分。空白对照也显色的原因极有可能是封闭不彻底!

我用的是含1%BSA的TBST封闭液,30min,这样不彻底吗,那如何改进呢
3楼2013-10-01 13:55:24
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yyc12596

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引用回帖:
3楼: Originally posted by xinyuecao123 at 2013-10-01 13:55:24
我用的是含1%BSA的TBST封闭液,30min,这样不彻底吗,那如何改进呢...

1%BSA in PBS, incubate at room temperature for a minimum of 1 hour.

      Advice: >= 1 hour. 30 mins is not enough.
4楼2013-10-09 02:45:33
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