Anti-Epo receptor antibodies do not predict Epo receptor expression
Anti-Epo receptor antibodies do not predict Epo receptor expression
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DOI：
10.1182/blood-2005-10-4066
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发表时间：
2006-03-01
期刊：
影响因子：
20.3
通讯作者：
Begley, CG
中科院分区：
文献类型：
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作者：
Elliott, S;Busse, L;Begley, CG
研究人员使用抗EpoR抗体进行免疫印迹和免疫染色，报告了促红细胞生成素受体（EpoR）在非造血组织，包括人类肿瘤中的表达。然而，这些抗体检测到66至78 kDa的蛋白质，显著大于EpoR的预测分子量（56-57 kDa）。我们研究了这些抗体的特异性，并表明它们都检测到非EpoR蛋白。C-20在肿瘤细胞系中检测到3种蛋白（35、66和100 kDa）。从制备凝胶中获得的序列与C-20免疫肽具有相似性。66-kDa蛋白是热休克蛋白（HSP 70），在肽竞争实验中抗体结合被废除。抗体M-20容易鉴定59-kDa EpoR蛋白。然而，无论是M-20还是C-20都不适合使用免疫组织化学方法检测EpoR。我们得出结论，这些抗体用于检测EpoR的效用有限。因此，使用这些抗体在肿瘤细胞中表达EpoR的报告应谨慎看待。
Investigators using anti-EpoR antibodies for immunoblotting and immunostaining have reported erythropoietin receptor (EpoR) expression in nonhematopoietic tissues including human tumors. However, these antibodies detected proteins of 66 to 78 kDa, significantly larger than the predicted molecular weight of EpoR (56-57 kDa). We investigated the specificity of these antibodies and showed that they all detected non-EpoR proteins. C-20 detected 3 proteins in tumor cell lines (35, 66, and 100 kDa). Sequences obtained from preparative gels had similarity to the C-20-immunizing peptide. The 66-kDa protein was a heat shock protein (HSP70) to which antibody binding was abrogated in peptide competition experiments. Antibody M-20 readily identified a 59-kDa EpoR protein. However, neither M-20 nor C-20 was suitable for detection of EpoR using immunohistochemical methods. We concluded that these antibodies have limited utility for detecting EpoR. Thus, reports of EpoR expression in tumor cells using these antibodies should be viewed with caution.