Brizzard B L, Chubet R G, Vizard D L
Eastman Kodak Company, New Haven, CT.
Biotechniques. 1994 Apr;16(4):730-5.
The FLAG epitope is an eight amino acid peptide (AspTyrLysAspAspAspAspLys) that is useful for immunoaffinity purification of fusion proteins. A monoclonal antibody (anti-FLAG M1) that binds the FLAG epitope in a calcium-dependent manner and requires an N-terminal FLAG sequence has been described previously. We describe the use of a second anti-FLAG monoclonal antibody (anti-FLAG M2) in immunoaffinity purification of N-terminal Met-FLAG and C-terminal FLAG fusion to bacterial alkaline phosphatase. Although binding of an anti-FLAG M2 monoclonal antibody to the FLAG epitope is not calcium-dependent, bound fusion proteins can be eluted by competition with FLAG peptide.
FLAG表位是一种八氨基酸肽（天冬氨酸-酪氨酸-赖氨酸-天冬氨酸-天冬氨酸-天冬氨酸-天冬氨酸-赖氨酸），可用于融合蛋白的免疫亲和纯化。之前已描述过一种单克隆抗体（抗FLAG M1），它以钙依赖方式结合FLAG表位，且需要N端的FLAG序列。我们描述了第二种抗FLAG单克隆抗体（抗FLAG M2）在免疫亲和纯化N端甲硫氨酸-FLAG和C端FLAG与细菌碱性磷酸酶融合蛋白中的应用。虽然抗FLAG M2单克隆抗体与FLAG表位的结合不依赖钙，但结合的融合蛋白可通过与FLAG肽竞争而洗脱。
