Monoclonal antibodies to two different epitopes in a 30-kD CNBr peptide of the K1 and K2 keratins.

Monoclonal antibodies to two different epitopes in a 30-kD CNBr peptide of the K1 and K2 keratins.
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针对 K1 和 K2 角蛋白的 30-kD CNBr 肽中两个不同表位的单克隆抗体。

DOI:
10.1111/1523-1747.ep12514321
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发表时间:
1990
期刊:
The Journal of investigative dermatology
影响因子:
--
通讯作者:
Goldsmith,LA
Goldsmith,LA
中科院分区:
--
文献类型:
--
作者:
Colbert,MC;McCoon,PE;Day,KH;Lane,AT;Goldsmith,LA

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制备了两种抗角蛋白单克隆抗体Kab-2和Kab-3,它们对II型(碱性)人表皮角蛋白的不同表位具有特异性。这些抗体在人胎儿表皮上有不同的免疫荧光染色模式。Western blots和固相RIA检测结果显示,这两种抗体均与从包皮表皮提取的65- 67-kD碱性角蛋白(K1和K2)结合。与两种Kab抗体和其他抗角蛋白单克隆抗体的竞争结合研究表明,Kab-2和Kab-3识别的相关表位与其他抗角蛋白抗体AE-1、2和3识别的表位不同,Kab-2和表位与金黄色葡萄球菌v8蛋白酶消化K1角蛋白产生的肽的反应性不同;抗体可以识别常见和独特的肽。溴化氰酶切K1角蛋白的Western blot结果显示,两种Kab抗体都与该分子的一个30-kD片段发生反应,该片段被认为是n端CNBr肽。我们解释这些数据表明,在组织中,K1角蛋白的n端区域部分与这些单克隆抗体的反应是不同的,并且对同一分子的单克隆抗体染色的形态学差异可以反映与超分子组织相关的表位特异性或表位可用性。
Two anti-keratin monoclonal antibodies, Kab-2 and Kab-3, with specificities for different epitopes of type II (basic) human epidermal keratins, were produced. These antibodies had different immunofluorescent staining patterns on human fetal epidermis. Western blots and solid phase RIA showed both antibodies bound to 65- 67-kD basic keratins (K1 and K2) extracted from foreskin epidermis. Competitive binding studies with the two Kab antibodies and other anti-keratin monoclonal antibodies showed that Kab-2 and Kab-3 recognized related epitopes, distinct from the epitopes recognized by other anti-keratin antibodies AE-1, 2, and 3 Kab-2 and epitopes were epitopes were distinguished by differences in their reactivity with peptides generated byStaphlococcus aureusV8 protease digestion of the K1 keratin; the antibodies recognized both common and unique peptides. Western blots of cyanogen bromide digests of the K1 keratin showed that both Kab antibodies reacted with a 30-kD fragment of the molecule presumed to be the N-terminal CNBr peptide. We interpret these data to indicate that in tissues, portions of the N-terminal region of the K1 keratin are differentially available for reaction with these monoclonal antibodies and that morphologic differences in staining with monoclonal antibodies to the same molecule can reflect epitope specificity or epitope availability related to supramolecular organization.
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