Protein Biotinylation Kit
For in vitro use only!
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Binding of biotin by avidin, streptavidin or NeutrAvidin™ is the strongest known biological interaction with a dissociation constant in the range of 10-15 M.
The vitamin biotin may be conjugated to many proteins without loss of their biological activity due to the small size of the biotin molecule. The biotinylated probe is usually detected by avidin, streptavidin or NeutrAvidin™, carrying a reporter group, e.g. horseradish peroxidase (HRP) or a fluorescent label.
The protein of interest (POI) is often biotinylated at different positions. Each conjugated biotin binds one molecule of avidin, streptavidin or NeutrAvidin™, resulting in signal amplification.
Biotin-X inserts a C6-spacer between the biotin and the protein molecule, hence reduces steric hindrance and increases accessibility for avidin, streptavidin or NeutrAvidin™.
The interaction between biotin and biotin-binding proteins is used in various applications such as affinity chromatography, fluorescence-activated cell sorting (FACS), ELISA and Western Blot.
This Protein Biotinylation Kit contains all reagents required for performing 5 separate labeling reactions of 5 mg of POI.
5 vials containing 1 mg each
1 vial containing 168 mg
Storage and Stability:
Upon receipt, store the biotin-X at -20 °C. The other components may be stored at room temperature. If stored as recommended, Jena Bioscience guarantees optimal performance of this product for 12 months.
The protein concentration should be at least 2 mg/ml, higher concentrations - up to 10 mg/ml - are preferable since labeling efficiency suffers from low concentrations. We recommend using about 5 mg protein per labeling reaction.
Buffers containing primary amines such as Tris and glycine are not suitable for the labeling reaction and must be exchanged with suitable amine-free buffer such as PBS, MES, or HEPES before starting the labeling reation.
Storage of the Conjugate
Store the conjugate just like the unlabeled protein. We recommend dividing the solution into small aliquots and freeze at -20 °C or -80 °C. Avoid repeated freezing and thawing!
Low biotinylation efficiency
Note that biotinylation efficiency not only depends on the surrounding conditions but also on the protein characteristics. The tertiary structure and the resulting number of lysines on the surface of the protein play a role as well as the isoelectrical point and the behaviour of the protein at pH 8.3.