Identification of Interacting Motifs Between Armadillo Repeat Containing 1 (ARC1) and Exocyst 70 A1 (Exo70A1) Proteins in Brassica oleracea;PROTEIN JOURNAL 卷: 35 期: 1 页: 34-43 出版年: FEB 2016
A Novel Polyclonal Antiserum against Toxoplasma gondii Sodium Hydrogen Exchanger 1;KOREAN JOURNAL OF PARASITOLOGY 卷: 54 期: 1 页: 21-29 出版年: FEB 2016
A pair of transposon‐derived proteins regulate active DNA demethylation in Arabidopsis;EMBO JOURNAL 卷: 35 期: 18 页: 2060-+ 出版年: SEP 15 2016
泛素结合酶RAD6多克隆抗体的制备及初步鉴定;中国处方药
Orf Virus 002 Protein Targets Ovine Protein S100A4 and Inhibits NF-κB Signaling;FRONTIERS IN MICROBIOLOGY 卷: 7 文献号: 1389 出版年: SEP 13 2016
N-terminal domains of ARC1 are essential for interaction with the N-terminal region of Exo70A1 in transducing self-incompatibility of Brassica oleracea;ACTA BIOCHIMICA ET BIOPHYSICA SINICA 卷: 48 期: 9 页: 777-787 出版年: SEP 2016
Identification of Interacting Motifs Between Armadillo Repeat Containing 1 (ARC1) and Exocyst 70 A1 (Exo70A1) Proteins in Brassica oleracea;PROTEIN JOURNAL 卷: 35 期: 1 页: 34-43 出版年: FEB 2016
The potassium channel FaTPK1 plays a critical role in fruit quality formation in strawberry (Fragaria× ananassa);Plant Biotechnology Journal (2017), pp. 1–12,DOI: 10.1111/pbi.12824
Functional characterization of rice CW-domain containing zinc finger proteins involved in histone recognition;Plant Science 263 (2017) 168–176,DOI:10.1016/j.plantsci.2017.06.013
a. 洗脱时间不足;
b. 增大Elution Buffer的洗脱体积;
c. 加大Elution Buffer中还原型谷胱甘肽的浓度。谷胱甘肽的推荐浓度是10 mM,若此浓度洗脱效果不是很理想可以尝试50 mM Tris- HCl, 20–40 mM reduced glutathione, pH 8.0 的Elution Buffer;
d. 增大Elution Buffer的pH值。Elution Buffer的pH值调至pH 8–9可以提高洗脱效率而不需要增加glutathione的浓度;
e. 增加Elution Buffer的离子强度。加入0.1–0.2 M NaCl能提高洗脱效率;
f. 改用新鲜配置的Elution Buffer;
g. Elution Buffer中加入非离子型变性剂。非特异性的疏水作用可能会阻碍GST融合蛋白从磁珠上增溶和洗脱。加入0.1% Triton X-100 or 2% 和N-octylglucoside可以显著增加洗脱效率。
GST融合蛋白不与磁珠结合
a. 超声太剧烈或时间过长会引起蛋白变性,导致蛋白不能与磁珠结合。需注意设置好超声仪的功率和间隔时间;
b. 在细胞裂解和加Binding Buffer和Elution Buffer之前,加入终浓度1–10 mM DTT可以显著提高GST融合蛋白的结合效率;
c. 由于pH值低于6.5或高于8.0结合效率会降低,因此使用前需用pH6.5~8.0的Buffer如PBS进行平衡;
d. 如果磁珠已经使用过几次,有必要进行重生或者改用新磁珠