当前位置： SCI文献检索 > JOURNAL OF PROTEOME RESEARCH期刊下所有文献 > Variability in Streptavidin-Sepharose Matrix Quality Can Significantly Affect Proximity-Dependent Biotinylation (BioID) Data.

Variability in Streptavidin-Sepharose Matrix Quality Can Significantly Affect Proximity-Dependent Biotinylation (BioID) Data.

Abstract：

:Due to their ease of use and high binding affinity, streptavidin-based purification tools have become widely used for isolating biotinylated compounds from complex mixtures. We and others routinely use streptavidin-sepharose matrices to isolate biotinylated polypeptides generated in proximity-dependent biotinylation approaches, such as BioID or APEX. However, we noted sporadic, substantial variation in the quality of BioID experiments performed in the same laboratories over time, using seemingly identical protocols. Identifying the source of this problem, here, we highlight considerable variability in streptavidin contamination derived from different production lots of streptavidin-sepharose beads from the same manufacturer and demonstrate that high levels of streptavidin peptide contamination can have detrimental effects on BioID data. We also describe two simple, rapid approaches to assess the degree of streptavidin "shedding" from individual lots of the sepharose matrix before use to avoid the use of lower quality reagent.

journal_name

J Proteome Res

journal_title

Journal of proteome research

authors

St-Germain JR,Samavarchi Tehrani P,Wong C,Larsen B,Gingras AC,Raught B

doi

10.1021/acs.jproteome.0c00117

subject

Has Abstract

pub_date

2020-08-07 00:00:00

pages

3554-3561

issue

8

eissn

1535-3893

issn

1535-3907

journal_volume

19

pub_type

杂志文章

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