Assessment of multiple different estrogen receptor-β antibodies for their ability to immunoprecipitate under chromatin immunoprecipitation conditions
Assessment of multiple different estrogen receptor-β antibodies for their ability to immunoprecipitate under chromatin immunoprecipitation conditions
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DOI：
10.1007/s10549-006-9229-5
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发表时间：
2006-05
影响因子：
3.8
通讯作者：
G. Weitsman;G. Skliris;K. Ung;B. Peng;M. Younes;P. Watson;L. Murphy
中科院分区：
文献类型：
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作者：
G. Weitsman;G. Skliris;K. Ung;B. Peng;M. Younes;P. Watson;L. Murphy
几种不同的总雌激素受体(ER)β、ERβ1和ERβ2/cx抗体在染色质免疫沉淀条件（ChIP）下免疫沉淀ERβ特异性亚型的能力进行了测试和比较。兔多克隆抗体AP-ERβ1和AP-ERβ2/cx分别特异于ERβ1和ERβ2/cx亚型，对ChIP最有效。单克隆抗体MCA1974/PPG5/10也能ChIP ERβ1，但效率低于AP-ERβ1。所有其他抗体不适合ChIP分析，尽管大多数抗体在标准条件下免疫沉淀出适当的ERβ亚型。为了鉴定也可用于验证体内表达谱的抗体，还进行了western blotting和免疫组织化学检测ERβ亚型的抗体的比较。在曼尼托巴乳腺肿瘤库的组织处理和自染色条件下，发现385P/GC17、MCA1974/PPG5/10、Ab288/14C8和MCA2279S/57/3对人乳腺组织活检切片中ERβ亚型的免疫组化效果最好，而Ab14021、AP-ERβ1和AP-ERβ2/cx对ERβ亚型的免疫印迹检测效果最好。
Several different antibodies to total estrogen receptor (ER)β, ERβ1 and ERβ2/cx have been tested and compared for their ability to immunoprecipitate ERβ specific isoforms under chromatin immunoprecipitation conditions (ChIP). The rabbit polyclonal antibodies AP-ERβ1 and AP-ERβ2/cx, specific for ERβ1 and ERβ2/cx isoforms, respectively, were the most efficient for ChIP. The monoclonal antibody MCA1974/PPG5/10 was also able to ChIP ERβ1, but less efficiently than AP-ERβ1. All other antibodies tested were not suitable for ChIP analyses although most antibodies tested immunoprecipitated the appropriate ERβ isoforms under standard conditions. To identify antibodies that can also be used to verify in-vivo expression profiles, a comparison of the antibodies to detect ERβ isoforms by western blotting and immunohistochemistry was also undertaken. Under the tissue processing and autostaining conditions used at the Manitoba Breast Tumor Bank 385P/GC17, MCA1974/PPG5/10, Ab288/14C8 and MCA2279S/57/3 were found to be the best for IHC of ERβ isoforms in human breast tissue biopsy sections, while Ab14021, AP-ERβ1 and AP-ERβ2/cx were best for western blot detection of ERβ isoforms.