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Description

Applications:

Biotin-Separopore® binds with high affinity to avidin and streptavidin and is used to purify (strept)avidin linked biomolecules such as proteins coupled to strept(avidin). The interaction of biotin with avidin is the strongest known non-covalent interaction between a protein and its ligand (Ka = 1015 M-1). Binding is rapid and is unaffected by most extremes of pH, organic solvents and other denaturing agents. Immobilized biotin can be released by exposure to 8M guanidine-HCl at pH 1.5 or if boiled in reducing SDS-PAGE sample Buffer. These elution conditions permanently denature the avidin protein. It has been widely used in biochemistry and molecular biology for detecting and labeling proteins. For mild conditions, use 2-Iminobiotin-Separopore® 4B-CL (SKU # 20181056).

Note: Separopore® is a cost-effective equivalent to Sepharose® in all of its physical properties and binding characteristics.

References

Development of the novel PEG-PE-based polymer for the reversible attachment of specific ligands to liposomes: synthesis and in vitro characterization. Bioconjug Chem. (2011) 22: 2005-13.
Single bead-based electrochemical biosensor. Biosens Bioelectron. (2009) 25: 809-14.
Quantification of specific bindings of biomolecules by magnetorelaxometry. J Nanobiotechnology. (2008) 6: 4.
Intracellular production of a soluble and functional monomeric streptavidin in Escherichia coli and its application for affinity purification of biotinylated proteins. Protein Expr Purif. (2006) 46: 268-73.
Antigen targeting to dendritic cells with bispecific antibodies. J Immunol Methods. (2005) 306: 80-92.
Engineering soluble monomeric streptavidin with reversible biotin binding capability. J Biol Chem. (2005) 280: 23225-31.
p-Nitrophenylcarbonyl-PEG-PE-liposomes: fast and simple attachment of specific ligands, including monoclonal antibodies, to distal ends of PEG chains via p-nitrophenylcarbonyl groups. Biochim Biophys Acta. (2001) 1511: 397-411.
Rapid purification of recombinant proteins fused to chicken avidin. Gene. (1995) 167: 63-8.
Affinity probes for the GABA-gated chloride channel: 5e-tert-butyl-2e-[4-(substituted-ethynyl)phenyl]-1,3-dithianes with photoactivatable, fluorescent, biotin, agarose and protein substituents. Bioorg Med Chem. (1995) 3: 1675-84.
Engineered chimeric streptavidin tetramers as novel tools for bioseparations and drug delivery. Biotechnology (N Y). (1995) 13: 1198-1204.
The iron-responsive element binding protein: a method for the affinity purification of a regulatory RNA-binding protein. Proc Natl Acad Sci U S A. (1989) 86: 5768-72.
Demonstration in children of oligoclonal IgG bands in unconcentrated CSF using agarose isoelectric focusing and immunolabeling. Pediatr Neurol. (1986) 2: 286-9.
Synthesis and use of the protein phosphatase affinity matrices microcystin-sepharose and microcystin-biotin-sepharose. Methods Mol Biol. (2007) 365: 39-45.
Characterization of the subunit structure of pyruvate carboxylase from Pseudomonas citronellolis. J Biol Chem. (1981) 256: 11819-25.

Properties

Shelf life

2 years

Storage Temperature

2-8°C

ECCN #

EAR99

Appearance color

White

Appearance form

Suspension

Matrix

Separopore® 6B-CL (crosslinked agarose beads, 6%)

Particle Size Range

53 - 180 μm

Matrix Activation

Carboxy

Matrix Attachment

Carboxy (amide linkage through diaminohexane spacer); matrix contains residual amino groups which are charged at physiological pH

Spacer Arm

16 atoms

Binding Capacity

~4 mg of avidin / ml drained gel

Suspension/Column/Cartridge

Supplied as suspension in Buffered saline containing 0.02% sodium azide

pH Stability

4 - 9

Flow Specifications

70 - 140 cm / h

Ligand Density

2 – 6 μmol / ml drained gel

Molecular Weight Range

6 x 104 - 2 x 107