Protein Purification
Baculovirus Expression
Mammalian Expression
E. coli Expression
Molecular Biology
Products
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Products: Eznyme Toolbox: TurboBiotinylation DT Kit
TurboBiotinylation DTTM Kit
| Catalog #: | Size: | Concentration: | Price: |
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| B0112P | 50X TurboBiotinylase DT (0.1 mg)
10X TurboBiotinylation Mix (1 ml) | 5 mg/ml | $360 |
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| B0112M | 50X TurboBiotinylase DT (1 mg)
10X TurboBiotinylation Mix (1 ml) | 5 mg/ml | $2,000 |
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| B0112L | 50X TurboBiotinylase DT (5 mg)
10X TurboBiotinylation Mix (5 ml) | 5 mg/ml | $6,200 |
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| T0112B | Bulk | 5 mg/ml | Inquiry |
Components 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 Purity:
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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:
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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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