Biotin Separopore® (Agarose) 4B-CL

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 (K_a = 10¹⁵ M^-1). Bond formation is rapid and is unaffected by most extremes of pH, organic solvents and other denaturing agents. Immobilized biotin will release avidin if exposed 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).

Technical specifications:

• Ligand: Biotin
• Matrix: Separopore 4B-CL (crosslinked agarose beads, 4%)
• Particle size range: 52 - 165 µm
• Molecular weight range: 6 x 10⁴ - 2 x 10⁷
• Matrix activation: Epoxy
• Matrix attachment: Carboxy (amide linkage through diaminopropane spacer); matrix contains residual amino groups which are charged at physiological pH
• Matrix spacer: 13 atoms
• Ligand density: 2 - 6 μmol / ml drained gel
• Binding capacity: 20 - 30 mg of avidin / ml drained gel
• pH stability: 4 - 9
• Flow specifications: 70 - 140 cm / h
• Supplied as suspension in phosphate buffered saline containing 0.02% sodium azide

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.

Related products:

• 2-Iminobiotin-Separopore 6B-CL (SKU # 20181056)

Usage : bioWORLD's products are supplied for LABORATORY RESEARCH USE ONLY. The product may not be used as a drug, agricultural or pesticidal product , food additive or as a household chemical.