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

Applications:

AMP Separopore interacts strongly with NAD⁺-dependent dehydrogenases and ATP-dependent enzymes. Selective elution with gradients of NAD⁺ or NADP⁺ allows the resolution of complex mixtures of dehydrogenase isoenzymes using 5’-AMP Separopore.

The production method involves alkylation of the nucleotide, AMP 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 Specifications:

• Ligand: Adenosine Monophosphate (AMP)
• Matrix: Separopore 4B-CL (highly crosslinked agarose beads, 4%).
• Particle size range: 52 - 165 µm
• Matrix activation: Epoxy
• Matrix attachment: N6
• Spacer arm: 1, 6-diaminohexane
• Ligand density: 2 µmol AMP / ml drained gel
• Binding capacity: ~ 5-7 mg lactate dehydrogenase / ml drained gel
• pH stability: 4 - 10
• Flow rate specifications: 70 - 140 cm / h.
• Chemical stability: Stable to all commonly used aqueous buffers and additives like detergents. Avoid high concentrations of EDTA, urea, guanidine-HCl, chaotropic salts and strong oxidizing agents
• Physical stability: Negligible volume variation due to changes in pH or ionic strength
• Supplied as lyophilized powder and 1g yields 8-10 ml of gel

References:

• Association of (c)AMP-degrading glycosylphosphatidylinositol-anchored proteins with lipid droplets is induced by palmitate, H₂O₂ and the sulfonylurea drug, glimepiride, in rat adipocytes. Biochemistry. (2008) 47: 1274-87.
• Active and inactive ecto-5'-nucleotidase variants in liver of control and dystrophic Lama2dy mice. Int J Biochem Cell Biol. (2004) 36: 422-33.
• Human aldehyde dehydrogenase 3A1 (ALDH3A1): biochemical characterization and immunohistochemical localization in the cornea. Biochem J. (2003) 376: 615-23.
• Purification and properties of betaine aldehyde dehydrogenase from Avena sativa. J Plant Res. (2003) 116: 133-40.
• Purification and properties of aminoaldehyde dehydrogenase from Avena sativa. J Plant Res. (2002) 115: 393-400.
• Introduction of a (poly)histidine tag in L-lactate dehydrogenase produces a mixture of active and inactive molecules. Anal Biochem. (2001) 295: 257-61.
• Characterisation of a homogeneous plant aminoaldehyde dehydrogenase. Biochim Biophys Acta. (2000) 1480: 329-41.
• Purification, characterization, and localization of an ATP diphosphohydrolase in porcine kidney. Am J Physiol Renal Physiol. (2000) 278: F978-88.
• Identification of an NADH-cytochrome b(5) reductase gene from an arachidonic acid-producing fungus, Mortierella alpine 1S-4, by sequencing of the encoding cDNA and heterologous expression in a fungus, Aspergillus oryzae. Appl Environ Microbiol. (1999) 65: 3873-9.
• Biochemical properties of 5'-nucleotidase from mouse skeletal muscle. Biochim Biophys Acta. (1998) 1386: 16-28.
• Fast purification of thioredoxin reductases and of thioredoxins with an unusual redox-active centre from anaerobic, amino-acid-utilizing bacteria. Microbiology. (1998) 144: 793-800.
• Purification and characterisation of NADH oxidase from Thermus aquaticus YT-1 and evidence that it functions in a peroxide-reduction system. Eur J Biochem. (1998) 251: 935-45.
• Human liver fatty aldehyde dehydrogenase: microsomal localization, purification, and biochemical characterization. Biochim Biophys Acta. (1997) 1335: 99-110.
• Cloning, sequencing and functional expression of dihydrolipoamide dehydrogenase from the human pathogen Trypanosoma cruzi. Eur J Biochem. (1997) 243: 739-47.
• C1q is a nucleotide binding protein and is responsible for the ability of clusterin preparations to promote immune complex formation. Biochim Biophys Acta. (1996) 1297: 159-66.
• A low-Km 5'-nucleotidase from rat brain cytosolic fraction: purification, kinetic properties, and description of regulation by a novel factor that increases sensitivity to inhibition by ATP and ADP. J Neurochem. (1996) 67: 795-804.
• Partial characterization of a new nucleotide binding glycoprotein of hepatocyte plasma membrane. Biochem Pharmacol. (1996) 51: 1269-76.
• Identification and tissue-specific expression of a NADH-dependent activity of dihydropyrimidine dehydrogenase in man. Anticancer Res. (1996) 16: 389-94.
• Purification and properties of human placental ATP diphosphohydrolase. Eur J Biochem. (1995) 234: 66-74.
• Preparation and characterization of isozymes and isoforms of horse liver alcohol dehydrogenase. J Chromatogr A. (1995) 711: 105-12.

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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.