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甲乙型流感病毒診斷試劑盒(愛思普林)
廣州健侖生物科技有限公司
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甲乙型流感病毒診斷試劑盒(愛思普林)
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【公司名稱】 廣州健侖生物科技有限公司
【市場部】 楊永漢
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【騰訊 】 2042552662
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號二期2幢101-103室
1、 一抗從4度拿出后,為什么要進(jìn)行37度復(fù)溫?
(1)一方面,防止切片從4度直接放入PBS易脫片;
(2)另一方面,使抗原抗體結(jié)合更穩(wěn)定。一般不需要,但對表達(dá)較弱的抗原可能有用,4度和37度時分子運(yùn)動方式不同,前者分子碰撞機(jī)率和運(yùn)動速度小于后者,后者結(jié)合更快,但敏感性也提高了并易造成非特異染色。
(3)其實(shí),我更贊同后一種說法,因?yàn)槲覈L試把肝臟或睪丸片子從4度過夜拿出后,直接用PBS洗沒發(fā)生過脫片現(xiàn)象。
2、切片染色后背景太深,如何區(qū)分特異性與非特異性著色?
全片著色是指整個切片全都染上了顏色,著色的強(qiáng)度可深可淺,總之,分不清那些組織是陽性那些組織是陰性。出現(xiàn)這種現(xiàn)象的原因有:
(1)抗體濃度過高:一抗?jié)舛冗^高是常見的原因之一。解決辦法是,每次使用新抗體前應(yīng)當(dāng)對其工作濃度進(jìn)行測試,使每一抗體個體化,找到適合自己實(shí)驗(yàn)室的理想工作濃度,既使是即用型的抗體也應(yīng)如此,不能只簡單的按說明書進(jìn)行染色。
(2)抗體孵育時間過長或溫度較高:解決辦法是,嚴(yán)格執(zhí)行操作規(guī)程,隨身佩帶報時表或報時鐘,及時提醒,避免因遺忘而造成時間延長?,F(xiàn)在流行的二步法(Polymer)敏感性很高,要求一抗孵育的時間不是傳統(tǒng)的1小時,而是30分鐘,因此,要根據(jù)染色結(jié)果進(jìn)行調(diào)整。
1, the first antibody out from 4 degrees, why 37 degrees rewarming?
(1) On the one hand, to prevent slicing directly into PBS from 4 degrees easy to strip;
(2) On the other hand, antigen-antibody binding is more stable. Generally not needed, but may be useful for poorly expressed antigens, 4 and 37 degrees when the molecular movement in different ways, the former molecular collision probability and speed less than the latter, the latter combination of faster, but the sensitivity also increased and easy Cause non-specific staining.
(3) In fact, I agree with the latter statement, because I tried to wash the liver or testis out of the night after 4 degrees, directly washed with PBS did not have the phenomenon of delamination.
2, slice background is too dark, how to distinguish between specific and non-specific coloring?
Whole-piece coloring refers to the entire section of the color dyed, the depth of the shading can be shallow, in short, can not l those organizations are positive those organizations are negative. The reasons for this phenomenon are:
(1) antibody concentration is too high: a high concentration of anti-one is a common cause. The solution is to test the working concentration before each use of the new antibody so that each antibody is individualized to find the ideal working concentration for your laboratory, even for ready-to-use antibodies, not just simple According to the instructions for staining.
(2) antibody incubation time is too long or high temperature: The solution is to strictly enforce the rules, it is best to wear timepieces or clock, timely reminder to avoid the delay caused by the forgotten. Now the popular two-step method (Polymer) high sensitivity, requiring a first antibody incubation time is not the traditional one hour, but 30 minutes, therefore, according to the staining results to be adjusted.