乙型流感病毒诊断试剂（美国OSOM）

乙型流感病毒诊断试剂（美国OSOM）

广州健仑生物科技有限公司

广州健仑长期供应各种流感检测试剂，包括进口和国产的品牌，主要包括日本富士瑞必欧、日本生研、美国BD、美国NovaBios、美国binaxNOW、日本积水、日本荣研、美国OSOM、芬兰、爱思普林、英国Clearview、凯必利、广州创仑等主流品牌。

乙型流感病毒诊断试剂（美国OSOM）

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【公司名称】 广州健仑生物科技有限公司
【市场部】    杨永汉

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【腾讯  】 2042552662
【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

非特异性染色
① 是否有效地去除了内源性酶和生物素。应注意的是，并不是每一种组织均需要进行此步骤，但对于内源性酶或生物素丰富的组织，如肝脏、肾脏等，需考虑此原因。处理的方法为：
灭活碱性磷酸酶：zui常用的方法是将左旋咪唑（24mg/m1）加入底物液中，并保持pH 值在7.6~8.2，即能除去大部分内源性碱性磷酸酶，对于仍能干扰染色的酸性磷酸酶，可用50mmol/L 的酒石酸抑制。
饱和处理内源性生物素：
消除内源性生物素的方法是事先滴加亲和素，以饱和内源性生物素，使之不再有剩余的结合位点。具体方法是在ABC 法或SP法染色前将切片浸于25ug/ml亲和素溶液中处理15 分钟，PBS 清洗15 分钟后即可染色。
② 是否选择使用了正确的封闭血清。电荷吸附所造成的非特异性背景染色消除方法是以二抗动物的非免疫血清，用PBS 稀释为3%-10%溶液孵育切片，以封闭吸附位点。有时其它无关蛋白，如牛血清白蛋白也常应用。另外，取材时避开出血、坏死区亦极重要。zui近有些国内的实验室应用5%脱脂奶粉替代血清进行抗原封闭，效果也不错。
③ 所选择的抗体是否符合试验要求。因抗原不纯、标本片中含有与靶抗原相似的抗原决定簇等原因造成的非特异性染色只能通过采用高纯度、高效价的抗体、或针对更具特异性抗原决定簇的单克隆抗体来解决。
④ 一抗的使用浓度是否过高。
⑤ 清洗是否充分。应严格操作规程。因在缓冲液中含有一定量的盐，这亦有利于减低背景着色，通常0.05mol/l Tris—HCl，0.15mol/l NaCl已适用于多数染色方法，溶液内加入吐温20，效果更佳。特殊标记时，试剂公司一般都提供缓冲液的配方。
⑥ DBA 的使用是否正确。DAB 的孵育时间和配制方式可以产生某些背景颜色，使用浓缩型DAB 试剂盒时，请严格按照说明书标明的滴加顺序操作，注意校正蒸馏水的pH 值，以确保实验结果的正确性；粉剂DAB 溶解时，常有一些不溶性颗粒，这些颗粒须经过滤除去，否则可能沉积于切片组织上，产生斑点状着色。另外，DAB 保存不妥产生氧化物亦可沉积于切片上，因此需将DAB 保存于避光干燥处，现用现配，临用前加H2O2。孵育时间过长也会造成背景染色。
⑦ 标本染色过程中是否曾经干涸，否则会造成边缘部的非特异性染色。
⑧ 检查二抗与标本的内源性组织蛋白是否有交叉反应。

Non-specific staining
① whether the effective removal of endogenous enzymes and biotin. It should be noted that this procedure is not required for every tissue, but for endogenous enzymes or biotin-rich tissues such as the liver, kidneys, etc., this reason needs to be considered. The method of treatment is:
Inactivation of alkaline phosphatase: The most commonly used method is to add levamisole (24 mg / ml) to the substrate and maintain pH between 7.6 and 8.2 to remove most of the endogenous alkaline phosphatase. For still Can interfere with staining of acid phosphatase, available 50mmol / L of tartaric acid inhibition.
Saturated treatment of endogenous biotin:
The method of eliminating endogenous biotin is to drip the avidin in advance to saturate the endogenous biotin so that it no longer has the remaining binding sites. The specific method is ABC method or SP staining prior to the slice immersed in 25ug / ml avidin solution for 15 minutes, PBS washed for 15 minutes after staining.
② choose to use the correct closed serum. Non-specific background staining caused by the charge-sorption method is based on the non-immune serum of the secondary antibody, incubated with 3% -10% diluted PBS solution to block the adsorption site. Sometimes other unrelated proteins, such as bovine serum albumin are often used. In addition, avoid bleeding when drawing, necrosis area is also very important. Recently some domestic laboratories apply 5% skim milk instead of serum for antigen blocking, the effect is not bad.
③ whether the selected antibodies meet the test requirements. Non-specific staining due to impure antigen, antigenic determinants in the specimen containing antigen similar to the target antigen can only be achieved by using high-purity, high-titer antibodies or monoclonal antibodies against more specific epitopes solve.
④ the use of the first antibody concentration is too high.
⑤ cleaning is adequate. Should be strictly operating procedures. It is also advantageous to reduce the background color because a certain amount of salt is contained in the buffer. Usually, 0.05 mol / l Tris-HCl and 0.15 mol / l NaCl are suitable for most staining methods. Tween 20 is added to the solution to give a better result . Reagents typically provide buffer solutions for special labeling.
⑥ DBA use is correct. DAB incubation time and preparation methods can produce some background color, the use of concentrated DAB kit, in strict accordance with the instructions specified in the order of dropwise addition, pay attention to correct the pH value of distilled water to ensure the correctness of the experimental results; DAB DAB When dissolved, there are often some insoluble particles, these particles have to be removed by filtration, or may be deposited on the tissue sections, resulting in spots colored. In addition, DAB improper storage of oxide can also be deposited on the slice, so the DAB should be stored in a dark place, is now available with, before the addition of H2O2. Long incubation time can also cause background staining.
⑦ specimens have been dried in the dyeing process, otherwise it will cause non-specific edge of the stain.
⑧ check the secondary antibodies and specimens of endogenous tissue proteins cross-reaction.