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IMINODIACETIC ACID-MATRIX

CHARACTERISTICS OF THE MATRIX

Matrix: SepharoseTM CL-4B
and/or SepharoseTM 4 Fast Flow
Activation method: Oxiran.
Binding capacity: 45 micrograms of Cu2+ atom-ion per ml of wet gel.
Stability of the matrix: pH 2-11.
Shelf life of the matrix: Several years, if stored at 4°C in the presence of NaN3 0.02% (w/v).
Life time of the matrix with routine use: Approx. 20 cycles with no loss of binding capacity.
Storage: 4°C in PBS pH 7.4; NaN3 0.02% (w/v).

ORDERING INFORMATION

Product code Product name Package *Price in EUR
IDA15-4S Iminodiacetic acid SepharoseTM
CL-4B
Matrix
100 ml 270.00
500 ml 930.00
1000 ml 1600.00
IDA15-4FF Iminodiacetic acid SepharoseTM
4 Fast Flow
Matrix
100 ml 400.00
500 ml 1400.00
1000 ml 2400.00
* All prices are FOB Liège.
Bulk quantities are available.

INSTRUCTION FOR USE (Metal Chelating)

  1. De-gas the ISA-matrix when using more than 25 ml of matrix.
  2. Pour the matrix into a glass column with a frit of porosity P.1.
  3. Wash the column with 10 volumes of distilled water.
  4. Apply a bivalent or trivalent metal ion solution such as Cu2+, Zn2+... at a concentration 5 mg/ml in distilled water, at a flow rate of 50 ml/cm2/hour, until saturation.
  5. Wash the column with distilled water until the O.D at 280 nm of the eluent falls to the baseline level.
  6. Equilibrate the column with the Tris 50mM, AcOH pH 8.0.
  7. The obtained metal chelate affinity matrix are ready-to-use.

Applications

  1. Zn(II) dependence of the Aeromonas hydrophyla AE036 Metallo beta-lactamase Activity and Stability; Maria Hernandez Valladares et al.; Biochemistry, 36, 11534-11541 (1997).
  2. Characterization and sequence of the Chryseobacterium (flavobacterium) meningosepticum carbapenemase: a new molecular class B beta-lactamase showing a broad substrate profile; Rossolini GM et al.; Biochem. J., 332, 145-152 (1998).
  3. Mono- and binuclear Zn(II) beta-lactamase, role of the conserved cystein in the catalytic mechanism; Raquel Paul-Soto et al.; J. Biol. Chem., 274, 13242-13249 (1999).
  4. Expression, purification, crystallization and preliminary X-ray analysis of Aeromonas hydrophilia metallo--lactamase Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 February 1; 61(Pt 2): 180182.

  PDC (pentadentate chelator) & PDC kit for IMAC
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