Start > Ausstellungen / Broschüren > Quarterly Printed Magazine "TCIMAIL" > Back Numbers > Pre-Weighed Biotinylation Reagents

Pre-Weighed Biotinylation Reagents

Detection and purification of proteins are the core methods in life science experiments. The biotin-avidin interaction is often used to detect or purify proteins because of its high binding affinity. 1 and 2 are biotinylation reagents containing both a linker and an N-hydroxysuccinimidyl ester (NHS) moiety and are pre-weighed in suitable amounts for protein biotinylation. They can biotinylate proteins by reacting with amino groups (-NH2) of proteins and by forming amide bonds at pH between 7 and 9. Below is the direction for use.
B6096,B6097
Preparation:
It is recommended to prepare a 10 mM biotinylation solution. In order to efficiently biotinylate a sample, biotinylation solution should be used at a 15-fold molar excess over the amount of amine-containing protein. Here shows the calculation of the 10 mM biotinylation solution amount.
Calculate Example: A μL of 10 mM biotinlation solution for biotinylation 2 mg IgG (150,000 M.W.)

2 [mg IgG] × 10-3 [g/mg] × 1/150,000 [mol/g] × 15 [fold]
= A [μL of 10 mM biotinylation solution] × 10-6 [L/μL] × 10 [mmol/L] × 10-3 [mol/mmol]

A = 20 [μL of 10 mM biotinylation solution]
Direction for Use
  1. Bring 1, 2 to room temperature and dissolve 2 mg of 1 in 350 μL of DMSO or DMF, or 2 mg of 2 in 400 μL of PBS to prepare a 10 mM biotinylation solution.
  2. Dissolve the sample (1-10 mg/mL) in an appropriate buffer with pH between 7 and 9 such as PBS. Do not use buffers including amines (such as Tris).
  3. Add A μL (calculated above) of 10 mM biotinylation solution to the sample solution and incubate the mixed solution for 30 min at room temperature.
  4. Remove unreacted and hydrolyzed reagent by a desalting column or dialysis.

References

  • D. Savage et al., in Avidin-Biotin Chemistry: A Handbook, 2nd ed., Pierce Chemical, 1992, pp. 42-43.
The prices are subject to change without notice. Please confirm the newest price by our online catalog before placing an order.
In addition, sales products changes with areas. Please understand that a product is not available when the product details page is not displayed.