TurboBiotinylation DT Kit-蚂蚁淘生物

货号：	B0112
保存条件：	4 oC
供应商：	上海经科化学科技有限公司
保质期：	All components of the kit can be stored at 4 oC for at least 6 months.
CAS号：	无
英文名：	TurboBiotinylation DT Kit
数量：	期货
规格：	50X TurboBiotinylase DT (0.1 mg)

TurboBiotinylation DT KitDescription:Accelagen\"s TurboBiotinylation DT^TM Kit is an enhanced biotinylation kit for in vitro biotinylation of biotin-accepting proteins and recombinant proteins with AviTag^TM or other biotin-accepting peptides. Combining concentrated TurboBiotinylase and stable 10X TurboBiotinylation Mix, this kit offers high efficiency in vitro biotinylation from 4 ^oC to 37 ^oC. It does not require any special buffer for its activity and is compatible with a wide range of buffers that are most suitable for target proteins. This kit works equally well across pH 5.5-10, between 0-1 M NaCl, and in 10% glycerol. Protein Type:biotin ligaseSource:E. coliSpecific Activity:TurboBiotinylase DT has a specific activity of >4,000 Units/µg.One unit of TurboBiotinylase DT completely biotinylates 1 pmol of a substrate protein (23 kDa) at concentration of 1 mg/ml within 1 hour at 25 ^oC.Storage and Handling:TurboBiotinylation DT Kit is shipped with gel packs.The kit retains its full activity after incubation at 25 ^oC for over a week.All components of the kit can be stored at 4 ^oC for at least 6 months.For long term storage, store 50X TurboBiotinylase DT at -80 ^oC and 10X TurboBiotinylation Mix at 4 ^oCComponents and Formulation:

50X TurboBiotinylase DT at 5 mg/ml (80 µM) in Storage Buffer of 50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 2 mM TCEP, 50% glycerol.
10X TurboBiotinylation Mix (50 mM Tris-HCl, pH 7.5, 100 mM MgCl₂, 100 mM ATP, 5 mM biotin)

TurboBiotinylase DT is a stable ultra-pure recombinant E. coli BirA biotin ligase with dual GST- and His-tags. It is purified from E. coli and has a calculated molecular weight of 62.4 kDa. It is free of protease or nuclease contamination. It can be easily removed by affinity capture through its GST- or His-tag.

The efficiency of biotinylation can be measured by mass spectrometry. Biotinylation adds 226.31 Da to a protein.

Alternatively, biotinylation can be evaluated by streptavidin-agarose pull-down. A Coomassie stained gel will visually show the efficiency of biotinylation.

Remove biotin and ATP from the biotinylation reaction sample by a spin desalting column.

Save sample as Total (T)

Incubate with streptavidin-agarose for 1 hour with agitation.

Spin down the streptavidin-agarose.

Save the supernatant as Unbound (U)

Save the streptavidin-agarose as Bound (B)

Run gel and stain with Coomassie Blue.

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