Use for the isolation and purification of biotinylated molecules. SoftLink™ Resin is a rigid, methacrylate polymeric gel filtration matrix, functionalized with covalently bound, monomeric avidin. Monomeric avidin binds biotin with a Kd value of 10–7M, allowing reversible binding of bound biotinylated proteins under mild elution conditions. Native, or tetrameric, avidin binds biotin with a very strong affinity (Kd = 10–15M), which in turn requires strong denaturing conditions for eluting bound material. Monomeric avidin allows the specificity of capture but also the mildness of release appropriate for the purification of sensitive biological materials.
SoftLink Resin binds 20–40nmol of biotinylated protein per milliliter of resin. Bound biotinylated molecules can be eluted under mild nondenaturing conditions (5mM biotin). The resin retains biotin binding ability following exposure to a wide range of pH, low or high ionic strength, 6M guanidine and 1% SDS, and can be regenerated at least 10 times without loss of binding capacity. SoftLink Resin supports high flow rates (300cm/hour) and centrifugal forces (1,500 × g) in batch applications and is usable in batch or column purification methods.
Applications
- Purification of biotinylated antibodies and other proteins.
- Purification of in vivo biotinylated fusion proteins from PinPoint™ Vectors.
- Reversible binding of biotinylated proteins under mild nondenaturing conditions.