Calmodulin Separopore® (Agarose) 4B

Applications:

Calmodulin-Separopore 4B is used for affinity purification of calmodulin-binding proteins including calmodulin-regulated proteins in eukaryotic cells and recombinant proteins containing calmodulin-binding peptide fusion tag. Calmodulin binds proteins principally through their interactions with hydrophobic sites on its surface. These sites are exposed after a conformational change induced by the action of Ca²⁺ on separate Ca²⁺-binding site. The binding of enzymes may be enhanced if the enzyme substrate is present and enzyme substrate calmoldulin-Ca²⁺ complexes are particularly stable.

Technical specifications

• Ligand: Calmodulin (Bovine)
• Matrix: Separopore 4B (agarose beads, 4 %)
• Particle size range: 52 - 165 µm
• Molecular weight range: 6 x 10⁴ - 2 x 10⁷
• Matrix activation: Cyanogen bromide
• Ligand density: ~ 1 mg calmodulin / ml drained gel
• Matrix attachment: Amino
• Matrix spacer: 1 atom
• pH stability: 4 - 9
• Storage: 2 - 8 C
• Chemical stability: Stable to all commonly used aqueous solutions
• Physical stability: Negligible volume variation due to changes in pH or ionic strength
• Supplied as suspension in 0.5M NaCl containing 0.02% thimerosal

References:

• Purification of Euphorbia characias latex peroxidase by calmodulin-affinity chromatography. Ital J Biochem. (2007) 56: 1-5.
• Ca²⁺/calmodulin-dependent cyclic nucleotide phosphodiesterase in cGMP metabolism in rabbit parotid acinar cells. Biomed Res. (2006) 27: 37-44.
• Use-dependent inhibition of the skeletal muscle ryanodine receptor by the suramin analogue NF676. Br J Pharmacol. (2005) 146: 525-33.
• Flagellar radial spokes contain a Ca²⁺-stimulated nucleoside diphosphate kinase. Mol Biol Cell. (2004) 15: 3891-902.
• Purification of a recombinant membrane protein tagged with a calmodulin-binding domain: properties of chimeras of the Escherichia coli nicotinamide nucleotide transhydrogenase and the C-terminus of human plasma membrane Ca²⁺-ATPase. Protein Expr Purif. (2004) 36: 31-9.
• Characterization of calmodulin binding to the orphan nuclear receptor Errgamma. Biol Chem. (2003) 384: 473-82.
• Calcium- and FK506-independent interaction between the immunophilin FKBP51 and calcineurin. J Cell Biochem. (2002) 84: 460-71.
• Ca²⁺ binding site 2 in calcineurin-B modulates calmodulin-dependent calcineurin phosphatase activity. Biochemistry. (2001) 40: 8808-14.
• Suramin and the suramin analogue NF307 discriminate among calmodulin-binding sites. Biochem J. (2001) 355: 827-33.
• Calmodulin enhances the stability of the estrogen receptor. J Biol Chem. (2001) 276: 17354-60.
• Cloning and expression of a cDNA encoding human inositol 1,4,5-trisphosphate 3-kinase C. Biochem J. (2000) 352: 343-51.
• Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs. glutamate at position 298. Proc Natl Acad Sci U S A. (2000) 97: 2832-5.
• The N-terminal region of the plasma membrane Ca²⁺ pump does not separate from the main catalytic fragments after proteolysis. Biochem Biophys Acta. (2000) 1464: 127-34.
• Nerve growth factor activation of the extracellular signal-regulated kinase pathway is modulated by Ca²⁺ and calmodulin. Mol Cell Biol. (2000) 20: 1931-46.
• Multiple forms of phosphatase from human brain: isolation and partial characterization of affi-gel blue binding phosphatases. Neurochem Res. (2000) 25: 107-20.
• Plasma membrane Ca²⁺ pump isoform 3f is weakly stimulated by calmodulin. J Biol Chem. (2000) 275: 4323-8.
• Calmodulin binds to p21(Cip1) and is involved in the regulation of its nuclear localization. J Biol Chem. (1999) 274: 24445-8.
• Calcium-calmodulin-dependent protein kinase II and protein kinase C-mediated phosphorylation and activation of D-myo-inositol 1,4, 5-trisphosphate 3-kinase B in astrocytes. J Biol Chem. (1999) 274: 14734-42.

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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.