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Description

Applications:

Ni2+-chelated Separopore® 4B-CL is an imminodiacetate (IDA)-immobilized metal affinity chromatography (IMAC) matrix that separates proteins via their differences in affinity to nickel metal ions. Ni2+-chelated Separopore® 4B-CL is ideal for purification of recombinant proteins containing (His)6 fusion tag.

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

Technical specifications:

  • Chelating group: Nickel
  • Metal ion capacity: >20µmol Ni2+ / ml drained gel

References

Overexpression of potassium channel ether à go-go in human osteosarcoma. Neoplasma. (2012) 59: 207-15.
Cloning and expression of a thermostable alpha-galactosidase from the thermophilic fungus Talaromyces emersonii in the methylotrophic yeast Pichia pastoris. J Microbiol Biotechnol. (2010) 20: 1653-63.
Antitumour response of a double mutant of staphylococcal enterotoxin C2 with the decreased affinity for MHC class II molecule. Scand J Immunol. (2010) 71: 169-75.
Polymorphic membrane proteins 1 and 7 from Chlamydophila felis are significant immunodominant proteins. Vet Microbiol. (2010) 144: 415-21.
Development of a bacteriophage-based system for the selection of structured peptides. Anal Biochem. (2009) 388: 122-7.
WNT5A is regulated by PAX2 and may be involved in blastemal predominant Wilms tumorigenesis. Neoplasia. (2008) 10: 1470-80.
Prokaryotic expression of truncated VP7 of bluetongue virus (BTV) and reactivity of the purified recombinant protein with all BTV type-specific sera. J Virol Methods. (2008) 152: 6-12.
Prophylactic evaluation of recombinant extracellular superoxide dismutase of Brugia malayi in jird model. Vaccine. (2008) 26: 3705-10.
A novel method to capture methylated human DNA from stool: implications for colorectal cancer screening. Clin Chem. (2007) 53: 1646-51.
The expression and purification of the N-terminal activation domain of the transcription factor c-Myc: a model substrate for exploring ERK2 docking interactions. Protein Expr Purif. (2007) 53: 80-6.
Recombinant single chain Fv antibodies specific for glycoprotein D of equid herpesvirus 1. Folia Microbiol (Praha). (2006) 51: 492-6.
FGF19 is a target for FOXC1 regulation in ciliary body-derived cells. Hum Mol Genet. (2006) 15: 3229-40.
Cloning of a gene fragment encoding bovine complement component C3d with expression and characterization of derived fusion proteins. Vet Immunol Immunopathol. (2006) 114: 61-71.
Production of recombinant Bartonella henselae 17-kDa protein for antibody-capture enzyme-linked immunosorbent assay. Diagn Microbiol Infect Dis. (2006) 55: 1-7.
16: Chiu J, Tillett D, March PE. Coexpression of the subunits of T7 DNA polymerase from an artificial operon allows one-step purification of active gp5/Trx complex. Protein Expr Purif. (2006) 47: 264-72.
Analysis of phosphatase and tensin homolog tumor suppressor interacting proteins by in vitro and in silico proteomics. Proteomics. (2005) 5: 1250-62.
Identification of target genes regulated by FOXC1 using nickel agarose-based chromatin enrichment. Invest Ophthalmol Vis Sci. (2004) 45: 3904-13.
Optimised expression and purification of recombinant human indoleamine 2,3-dioxygenase. Protein Expr Purif. (2004) 37: 392-8.
Prevention of aggregation after refolding by balanced stabilization-destabilization: production of the Arabidopsis thaliana protein APG8a (At4g21980) for NMR structure determination. Protein Expr Purif. (2004) 34: 280-3.
Expression and purification of turkey coronavirus nucleocapsid protein in Escherichia coli. J Virol Methods. (2004) 116: 161-7.
Heat elution chromatography of immunoglobulins. Protein Expr Purif. (2003) 30: 301-3.
Trichomonas vaginalis Hmp35, a putative pore-forming hydrogenosomal membrane protein, can form a complex in yeast mitochondria. J Biol Chem. (2003) 278: 30548-61.
Recombinant (2-->3)-alpha-sialyltransferase immobilized on nickel-agarose for preparative synthesis of sialyl Lewis(x) and Lewis(a) precursor oligosaccharides. Carbohydr Res. (2003) 338: 1153-61.
Expression and purification of a recombinant form of human aromatase from Escherichia coli. Biochem Pharmacol. (2002) 64: 1317-24.
Overexpression of Arabidopsis thaliana soluble epoxide hydrolase 1 in Pichia pastoris and characterisation of the recombinant enzyme. Protein Expr Purif. (2002) 26: 65-70.

Properties

Shelf life

2 years

Storage Temperature

2-8°C

ECCN #

EAR99

Hazmat Ship

Non-hazardous

Matrix

Separopore® 4B-CL (crosslinked agarose beads, 4%)

Particle Size Range

52 - 165 μm

Matrix Activation

IDA

Binding Capacity

~5 - 8 mg (His)6-tagged protein / ml medium

Suspension/Column/Cartridge

Supplied as suspension in 20% ethanol

pH Stability

2 - 11

Chemical Stability

For one week at 40 °C: 0.01M HCl, 0.1M NaOH, For 12 h: 1M NaOH, 70% acetic acid. For 30 min tested: 30% 2-propanol. For 1 h tested: 2% SDS

Binding Wash/Buffer

Stable in all commonly used buffers, reducing agents, denaturants, and detergents.