NHS-Activated Separopore® (Agarose) 4B-CL

Applications:

• Rapid immobilization of antibodies and other proteins to the matrix.
• Immobilization of ligands through primary amines to purify recombinant proteins.
• Immobilization of Protein A or Protein G for monoclonal antibody purification.
• Pre-activated medium for immobilization of ligands containing a primary amino group.
• N-hydroxyl succinimide (NHS) ester forms a stable amide (peptide) bond with the primary amines of ligands.
• The stability of an amide bond at high pH (up to pH 13) allows for a very stringent wash protocol during affinity purification using NHS-activated ligand coupling.
• Coupling can be carried out in neutral pH which makes it ideal for immobilization of antibodies, antigens, and other proteins sensitive to pH extremes.
• Coupling reaction can also be carried out in organic solvents.
• Provides a 10 atom spacer arm upon ligand coupling.
• Coupling completed after 2 - 4 h at room temperature.
• NHS-Activated Separopore provides a valuable tool for affinity purification of antibodies, antigens and other biomolecules.

Technical specifications:

• Group to be coupled: Primary -NH₂
• Matrix: Separopore 4B-CL (highly crosslinked agarose beads, 4%)
• Particle size range: 52 - 165 μm
• Molecular weight range: 6 x 10⁴ - 2 x 10⁷
• Chemical linkage of ligand: Amide bond
• Spacer arm: 6-aminocaproic acid, 10 atoms
• Substitution: >18 μmol NHS / ml drained gel
• pH stability: 3 - 13 (ligand dependent)
• Coupling conditions: pH 6 - 9; Temp: 4 - 25 C
• Storage: 2 - 8 C
• Flow rate specifications: 70 - 140 cm / h
• Supplied as a pre-swelled gel suspension in 100% isopropanol to protect the active groups

References:

• Epitope structure of the carbohydrate recognition domain of asialoglycoprotein receptor to a monoclonal antibody revealed by high-resolution proteolytic excision mass spectrometry. J Am Soc Mass Spectrom. (2011) 22: 148-57.
• Hepatitis B virus core interacts with the host cell nucleolar protein, nucleophosmin 1. J Microbiol. (2009) 47: 746-52.
• Using liposomal fluorescent biolabels to develop an immunoaffinity chromatographic biosensing system for biotin. Anal Chem. (2008) 80: 6405-9.
• Separation of protein C from Cohn Fraction IV-1 by mini-antibody. Adv Exp Med Biol. (2007) 599: 125-31.
• Chromatographic refolding of recombinant human interferon gamma by an immobilized sht GroEL191-345 column. J Chromatogr A. (2006) 1107: 192-7.
• Sephadex-based cell-affinity adsorbents: preparation and performance. Biotechnol Appl Biochem. (2002) 35: 55-60.
• Purification and characterization of cytokine-inducing protein of seed extract from Aeginetia indica L., a parasitic plant. Immunopharmacology. (2000) 49: 377-89.
• A method for the purification of cAMP-dependent protein kinase using immunoaffinity chromatography. Protein Expr Purif. (1998) 14: 418-24.
• Peptide synthesis on Sepharose beads. J Pept Res. (1997) 49: 355-62.
• Inhibition of Holliday structure resolving endonuclease VII of bacteriophage T4 by recombination enzymes UvsX and UvsY. J Mol Biol. (1997) 267: 150-62.
• Construction of a bioreactor to produce special breakdown products of phytate. J Biotechnol. (1996) 48: 153-9.
• Protein-protein interaction affinity chromatography of leukotriene C4 synthase. Protein Expr Purif. (1995) 6: 352-6.
• Recombinant human insulin receptor substrate-1 protein. Tyrosine phosphorylation and in vitro binding of insulin receptor kinase. J Biol Chem. (1995) 270: 4870-4.

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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.