Biotin blocking System
How to block non specific biotin ?
Adding BBS to the cell culture irreversibly binds free biotin, preventing it from breaking the avidin biotinylated receptor complexes. This allows for a 1-column 3 steps affinity purification process on agarose-avidin chromatography columns.
Step 0
Introduced in slight excess to the cell culture (5-10%), the Biotin Blocking System irreversibly binds free biotin. A standard HABA test can easily verify the results.
Step 1
The avidin-agarose chromatography column is activated with the ligandspecific biotinylated receptors (antibody-specific biotinylated antigens for example).
Step 2
The Biotin blocking System saturated culture medium is passed through and rinsed with an adapted running buffer, leaving the specific ligand-receptors immobilized. no interaction between the biotin from the cell culture with the avidin-agarose chromatography column. All ligand-specific biotinlated receptor remains in place for the binding with the ligand of interest.
Step 3
The ligand of interest is then eluted out of the affinity column with the appropriate releasing buffer.
1-column 3 steps process for cell culture media purification. Biotin Blocking System serves as safeguard for column integrity and performance.
Specifications
| Physicochemistry | BBS |
|---|---|
| Activity (U/ml) | > 70 |
| Avidin
(g/l) | 5.5-6.0 |
| Dry Matter
% (w:w) | < 11.0 |
| Salts (KCl/NaCl)
% (w:w) | < 5.0 |
| Sodium Azide
% (w:w) | 0.02 |
| Transmittance
OD (650nm) | < 0.1 |
| Colour | Yellow-reddish |
| Shelf life Years | 3 |
| Microbiology | BBS |
|---|---|
| Total Bacterial count in 1 mg | <1 |
| Other | BBS |
|---|---|
| Definition of activity | One Unit activity binds 1 µg of biotin |
| Country of Manufacturing | Belgium |
| Origin | Egg proteins from hens |