ATP Separopore® (Agarose) 4B-CL (Lyophilized)

Applications:

Adenosine 5'-triphosphate (ATP)-Separopore 4B-CL is used in the purification of ATP binding proteins. Any protein with an accessible ATP-binding domain will bind to ATP- Separopore 4B-CL, regardless of activation state. The activated proteins will bind to ATP-Separopore 4B-CL with higher affinity than the corresponding inactivated protein. This is important when purifying proteins from crude fractions where protein concentration is low, as this prevents non-specific binding. Better recovery and yields may be obtained when the ATP-Separopore 4B-CL purification is followed by ion exchange chromatography.

The production method involves alkylation of the nucleotide, ATP followed by alkaline rearrangement to yield the corresponding N6-carboxymethyl derivative with subsequent condensation using 1,6-diaminohexane to give N6-[(6-aminohexyl)carbamoylmethyl)].

Technical specification:

• Ligand: ATP
• Matrix: Separapore 4B-CL (crosslinked agarose beads, 4 %)
• Particle size range: 52 - 165 μm
• Matrix activation: Cyanogen bromide
• Ligand density: 2 - 5 ATP μmol / ml drained gel
• Attachment: N6 of the purine ring
• Spacer: 11 atoms
• Supplied as lyophilized powder, 1g powder yields 8 - 10 ml of gel

References:

• Chemoproteomic characterization of protein kinase inhibitors using immobilized ATP. Methods Mol Biol. (2012) 795: 119-34.
• Heme-dependent activation of neuronal nitric oxide synthase by cytosol is due to an Hsp70-dependent, thioredoxin-mediated thiol-disulfide interchange in the heme/substrate binding cleft. Biochemistry. (2011) 50: 7146-56.
• 6-Alkylsalicylic acid analogues inhibit in vitro ATPase activity of heat shock protein 90. Arch Pharm Res. (2010) 33: 1997-2001.
• Dominant-negative Hsp90 reduces VEGF-stimulated nitric oxide release and migration in endothelial cells. Arterioscler Thromb Vasc Biol. (2008) 28: 105-11.
• Structural basis for depletion of heat shock protein 90 client proteins by deguelin. J Natl Cancer Inst. (2007) 99: 949-61.
• Improved performance of column chromatography by arginine: dye-affinity chromatography. Protein Expr Purif. (2007) 52: 410-4.
• Unusual properties of the prespore-specific enzyme, UDPgalactose:polysaccharide galactosyl transferase, of Dictyostelium discoideum. J Basic Microbiol. (2004) 44: 459-70.
• Discovery of novel targets of quinoline drugs in the human purine binding proteome. Mol Pharmacol. (2002) 62: 1364-72.
• Characterization of the 105-kDa molecular chaperone. Identification, biochemical properties, and localization. Eur J Biochem. (2002) 269: 5632-41.
• Selected BTB/POZ-kelch proteins bind ATP. FEBS Lett. (2002) 516: 20-6.
• Cloning, sequencing, expression, and purification of the C isozyme of mouse phosphofructokinase. Protein Expr Purif. (1999) 16: 448-53.
• Secretory immunoglobulin A from human milk catalyzes milk protein phosphorylation. Appl Biochem Biotechnol. (1998) 75: 77-91.
• Identification and characterization of individual cyclin-dependent kinase complexes from Saccharomyces cerevisiae. Yeast. (1999) 15: 295-309.
• Molecular shape and ATP binding activity of rat p50, a putative mammalian homologue of RuvB DNA helicase. J Biochem. (1999) 125: 487-94.
• Rapid identification of protein phosphatase 1-binding proteins by mixed peptide sequencing and data base searching. Characterization of a novel holoenzymic form of protein phosphatase 1. J Biol Chem. (1998) 273: 24396-405.
• A nucleotide-binding domain of porcine liver annexin VI. Proteolysis of annexin VI labelled with 8-azido-ATP, purification by affinity chromatography on ATP-agarose, and fluorescence studies. Mol Cell Biochem. (1998) 181: 11-20.
• Rapid purification and characterization of nucleoside diphosphate kinase isoforms using ATP-sepharose affinity column chromatography. Mol Cells. (1997) 7: 630-4.
• Novel 30 kDa protein possessing ATP-binding and chaperone activities. Biochem J. (1997) 326: 567-72.
• Interaction of annexins IV and VI with ATP. An alternative mechanism by which a cellular function of these calcium- and membrane-binding proteins is regulated. FEBS Lett. (1997) 409: 300-6.
• Phosphorylation of different human milk proteins by human catalytic secretory immunoglobulin A. Biochem Mol Biol Int. (1996) 39: 521-7.

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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.