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Products: Eznyme Toolbox: TurboBiotinylation DT Kit

TurboBiotinylation DTTM Kit

Catalog #: Size: Concentration: Price:
B0112P 50X TurboBiotinylase DT (0.1 mg)
10X TurboBiotinylation Mix (1 ml)
5 mg/ml $285
B0112M 50X TurboBiotinylase DT (1 mg)
10X TurboBiotinylation Mix (1 ml)
5 mg/ml $1,625
B0112L 50X TurboBiotinylase DT (5 mg)
10X TurboBiotinylation Mix (5 ml)
5 mg/ml $5,400
T0112B Bulk 5 mg/ml Inquiry


To Order: Please email PO to order@accelagen.com or fax PO to 888-239-6490


PRODUCT DETAIL DATA SHEET PROTOCOL
Components and Formulation:
  1. 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.
  2. 10X TurboBiotinylation Mix (50 mM Tris-HCl, pH 7.5, 100 mM MgCl2, 100 mM ATP, 5 mM biotin)


TurboBiotinylase Purity:

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.


TurboBiotinylase Activity:

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.

  1. Remove biotin and ATP from the biotinylation reaction sample by a spin desalting column.
  2. Save sample as Total (T)
  3. Incubate with streptavidin-agarose for 1 hour with agitation.
  4. Spin down the streptavidin-agarose.
  5. Save the supernatant as Unbound (U)
  6. Save the streptavidin-agarose as Bound (B)
  7. Run gel and stain with Coomassie Blue.



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