Two embryonic, tissue-specific molecules identified by a double-label immunofluorescence technique for monoclonal antibodies.
Two embryonic, tissue-specific molecules identified by a double-label immunofluorescence technique for monoclonal antibodies.
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DOI：
10.1177/34.6.3084626
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发表时间：
1986-06
期刊：
影响因子：
--
通讯作者：
G. Wessel;D. McClay
中科院分区：
文献类型：
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作者：
G. Wessel;D. McClay
我们确定了两个组织特异性分子的海胆胚胎的免疫荧光技术能够共同定位单克隆抗体在同一组织切片。该技术使用单价Fab-荧光染料缀合物作为第二试剂，以避免后续抗体探针的串扰。使用这种技术，我们表明，两种细胞表面分子表达的不同细胞群在胚胎中。该技术通常适用于抗体，无论种属或亚型特异性如何，并使用市售试剂。该技术为分析工作提供了足够的扩增和分辨率，但对于筛选程序来说足够快。作为荧光复染剂，在方案中使用染料4-乙酰氨基-4 '-异硫氰酸基芪-2，2'-二磺酸（SITS）可提供组织的独特荧光背景染色，而不干扰特异性抗体染色。
We identify two tissue-specific molecules in the sea urchin embryo by an immunofluorescence technique capable of co-localizing monoclonal antibodies on the same tissue section. The technique uses monovalent Fab-fluorochrome conjugates as secondary reagents to avoid cross-talk of subsequent antibody probes. Using this technique, we show that two cell surface molecules are expressed by different cell populations in the embryo. The technique is generally applicable for antibodies regardless of species or subtype specificity, and uses commercially available reagents. The technique provides sufficient amplification and resolution for analytical work, yet is rapid enough for screening procedures. As a fluorescent counterstain, use of the dye 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) in the protocol provides a distinct fluorescent background staining of the tissue without interference with the specific antibody staining.