Detection of microgram quantities of carrier ampholytes in electrofocused proteins by thin-layer chromatography.
Detection of microgram quantities of carrier ampholytes in electrofocused proteins by thin-layer chromatography.
复制标题
通过薄层色谱法检测电聚焦蛋白质中微克量的载体两性电解质。
DOI：
10.1016/0003-2697(81)90046-4
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发表时间：
1981
影响因子：
2.9
通讯作者：
Watson,DW
中科院分区：
文献类型：
--
作者：
Bloomster,TG;Watson,DW
A rapid and sensitive method for the detection of carrier ampholyte contamination in electrofocused proteins is described. Samples containing proteins and carrier ampholytes were applied to cellulose thin-layer chromatographic sheets and developed in 10% trichloroacetic acid. Proteins and large-molecular-weight carrier ampholytes were precipitated at the origin while 10% trichloroacetic acid-soluble carrier ampholytes migrated as a diffuse ninhydrin (nitrogen)-positive area at an Rfgreater than 0.50. We found that 1.25 μg of carrier ampholytes contained enough 10% trichloroacetic acid-soluble components to be detected by thinlayer chromatography. Using this assay, we investigated techniques designed to remove carrier ampholytes from an electrofocused protein. Removal of large-molecular-weight components from carrier ampholytes by dialysis through a 3500 Mrcutoff membrane did not facilitate separation of carrier ampholytes from streptococcal pyrogenic exotoxin type C by dialysis or gel chromatography. Also, this protein binds irreversibly to mixed-bed ion-exchange resin. The best method for separating carrier ampholytes from streptococcal pyrogenic exotoxin type C was by electrodialysis at pH 4.0. Following electrodialysis, estimated carrier ampholyte contamination in this protein was less than 1 part in 500 parts (by weight).
