Guinea pig anti-zebrafish Rad21l1 polyclonal antibody production: An N-terminal fragment of Rad21l1 cDNA was amplified with Phusion DNA polymerase (Thermo Fisher Scientific, Catalog #: M0530L) using the following primers: Fwd: 5’-aactttaagaaggagatataccatgTCAAGCTTTTGCCTTCCTGT-3’ and Rev: 5’-tctcagtggtggtggtggtggtgctcAAGCATGCAGAAAAATAAGGCT-3’. The Rad21l1 PCR product was then cloned into pET28b using NEBuilder HiFi DNA Assembly Master Mix (NEB, Catalog #: E5520S). BL21 (DE3) cells containing pRARE and Rad21l1 overexpression construct were grown in 2.6 L of LB with kanamycin and chloramphenicol until an OD600 = 1 and induced with a final concentration of 1 mM IPTG at room temperature for six hours. The Rad21l1 peptide was purified under denaturing conditions using Novagen NiNTA purification resins (Sigma, Catalog #: 70666) according to the manufacturer’s instructions. The Rad21l1 peptide was concentrated to a final concentration of 1 mg/ml in PBS using a 10 kDa centrifugal filter (Sigma, Catalog # UFC901008). The Rad21l1-derived peptide was injected into three guinea pigs by Pocono Rabbit Farm and Laboratory following the 91-day polyclonal antibody production protocol.

注意:以上内容是从某篇研究文章中自动提取的,可能无法正确显示。



Q&A
请登录并在线提交您的问题
您的问题将发布在Bio-101网站上。我们会将您的问题发送给本研究方案的作者和具有相关研究经验的Bio-protocol成员。我们将通过您的Bio-protocol帐户绑定邮箱进行消息通知。