NHS-Activated Separopore® 4B-CL (With Spacer Arm)

NHS-Activated Separopore® 4B-CL (With Spacer Arm) aids with the rapid immobilization of antibodies and other proteins to the matrix. N-hydroxyl succinimide (NHS) ester forms a stable amide (peptide) bond with the primary amines of ligands. NHS-Activated Separopore® provides a valuable tool for affinity purification of antibodies, antigens and other biomolecules.

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

Application

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.

References

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.