Characterization of initiation factor 3 from wheat germ. 2. Effects of polyclonal and monoclonal antibodies on activity.
Characterization of initiation factor 3 from wheat germ. 2. Effects of polyclonal and monoclonal antibodies on activity.
复制标题
小麦胚芽起始因子 3 的表征。
DOI：
10.1021/bi00333a017
复制
发表时间：
1985
期刊：
影响因子：
2.9
通讯作者：
Ravel,JM
中科院分区：
文献类型：
--
作者：
Lauer,SJ;Browning,KS;Ravel,JM
材料和方法材料。 eIF-3 的制备方法如前一篇论文（Lauer 等人，1985）中所述。与辣根过氧化物酶缀合的山羊抗兔 IgG (H+ L) 和与辣根过氧化物酶缀合的兔抗小鼠 IgM+ IgA+ IgG、MonoAb Id EIA 试剂盒和 ABTS 获自 Zymed。硝化纤维试纸来自 Schleicher & Schuell，HRP 显色试剂来自 Bio-Rad Labora-tories.Assays。 eIF-3 支持多肽合成（测定 A）、结合 40S 核糖体亚基（测定 B）以及支持 125I 标记的 STNV RNA 与 40S 核糖体亚基结合（测定 C）的能力按照前述论文（Lauer 等，1985）中所述进行测定。 eIF-3 多克隆抗体的生产。一只 5 磅重的雄性新西兰兔肌肉注射了 0.2 毫克 eIF-3（CM-150 组分）和 0.7 毫升弗氏完全佐剂。两次加强注射，每次含 0.5 毫克
Materials and MethodsMaterials. eIF-3 was prepared as described in the preceding paper (Lauer et al., 1985). Goat anti-rabbit IgG (H+ L) conjugated to horseradish peroxidase and rabbit anti-mouse IgM+ IgA+ IgG conjugated to horseradish peroxidase, the MonoAb Id EIA kit, and ABTS were obtained from Zymed. Nitrocellulose paper was from Schleicher & Schuell, and the HRP color development reagent was from Bio-Rad Labora-tories.Assays. The abilities of eIF-3 to supportpolypeptide synthesis (assay A), to bind to 40S ribosomal subunits (assay B), and to support the binding of 125I-labeled STNV RNA to 40S ribosomal subunits (assay C) were determined as described in the preceding paper (Lauer et al., 1985). Production of Polyclonal Antibodies to eIF-3. A 5-lb, male New Zealand rabbit was injected intramuscularly with 0.2 mg of eIF-3 (CM-150 fraction) in 0.7 mL of Freund’s complete adjuvant. Two booster injections, each containing 0.5 mg of