A comparison of anti-biotin and biotinylated anti-avidin double-bridge and biotinylated tyramide immunohistochemical amplification.
A comparison of anti-biotin and biotinylated anti-avidin double-bridge and biotinylated tyramide immunohistochemical amplification.
复制标题
抗生物素和生物素化抗亲和素双桥和生物素化酪酰胺免疫组织化学扩增的比较。
DOI：
10.1016/s0165-0270(01)00454-x
复制
发表时间：
2001
影响因子：
3
通讯作者：
Maddox,MT
中科院分区：
文献类型：
--
作者：
Freedman,LJ;Maddox,MT
通常，用常规的免疫组织化学技术很难检测到非常少量的抗原。我们评估了三种扩增技术，包括抗生物素或抗亲和素双桥或生物素化酪胺扩增，以提高5-羟色胺转运体免疫组织化学的敏感性。对于抗生物素双桥，在二抗后，将切片在抗生物素抗体中孵育，然后在二抗中二次孵育，然后进行抗生物素-生物素-过氧化物酶复合体(ABC)的孵育。对于生物素标记的抗亲和素技术，在ABC之后，切片在生物素化的抗亲和素中孵育，然后在ABC中孵育。对于生物素化的酪胺技术，在ABC步骤之后，切片在生物素化的酪胺和过氧化氢中孵化，然后在ABC中孵化。抗生物素双桥还导致在不增加背景的情况下，染色纤维的数量和标记强度大幅增加。生物素标记的抗亲和素双桥也产生了显著的信号扩增，但背景显著。生物素化酪胺技术导致标记纤维的数量和染色强度的增加更大，背景染色适中。然而，这种优势在一次抗体的高稀释度时并不存在。因此，抗生物素双桥可能在免疫组织化学和免疫荧光以及其他需要提高生物素标志物的灵敏度和低背景的情况下有用。在某种程度的背景标记是可接受的情况下，生物素化酪胺技术也可能是有用的。
Often it is difficult to detect very small amounts of antigen with conventional immunohistochemical techniques. We evaluate three amplification techniques involving anti-biotin or anti-avidin double-bridges or biotinylated tyramide amplification to enhance the sensitivity of serotonin transporter immunohistochemistry. For the anti-biotin double-bridge, after the secondary antibody, the sections were incubated in anti-biotin antibody followed by a second incubation in the secondary antibody and then avidin–biotin–peroxidase complex (ABC). For the biotinylated anti-avidin technique, after the ABC, sections were incubated in biotinylated anti-avidin, followed by another incubation in ABC. For the biotinylated tyramide technique, after the ABC step, sections were incubated in biotinylated tyramide and hydrogen peroxide, followed by another incubation in ABC. The anti-biotin double-bridge also resulted in a large increase in the number of stained fibers and the intensity of labeling with no increase in background. A biotinylated anti-avidin double-bridge also produced significant signal amplification but significant background. The biotinylated tyramide technique resulted in an even larger increase in the number of labeled fibers and an intensity of their staining with a moderate amount of background staining. However, this advantage was not present at high dilutions of primary antibody. Thus, the anti-biotin double-bridge is likely to be useful in immunohistochemistry and immunofluorescence as well as other situations where increased sensitivity and low background from biotin markers is needed. The biotinylated tyramide technique may also be useful where some degree of background labeling is acceptable.