Certificate of Analysis and Data Sheet
Source:
E.Coli
Catalog No.
PRO-360
Background:
PEX can block angiogenesis and tumor growth in vivo, providing a potentially novel therapeutic approach for diseases associated with neovascularization. The appearance of PEX at sites of neovascularization may not only control normal angiogenesis, but when administered in sufficient quantities, may provide a naturally-occurring therapeutic inhibitor of diseases associated with angiogenesis.
Description :
Recombinant Human PEX (C-Terminal Fragment of Human Matrix Metalloproteinase-2, MMP-2) which comprises C-terminal hemopexin-like domain (amino acids 445-635), Recombinant Human PEX is a noncatalytic Metalloproteinase Fragment with Integrin Binding Activity and can inhibit cell associated collagenolytic activity both in vitro and in vivo.
PEX is expressed as inclusion body in E. coli and subsequently refolded in vitro to get biological activity.
Recombinant PEX is purified by proprietary chromatographic techniques.
Physical Appearance:
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation:
The protein (1mg/ml) was lyophilized with 2mM Tris pH-7.4.
Solubility:
It is recommended to reconstitute the lyophilized PEX in sterile 18MΩ-cm
H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.
Stability:
Lyophilized PEX although stable at room temperature for 3 weeks, should be stored desiccated
below -18 C. Upon reconstitution PEX should be stored at 4 C between 2-7 days and for future use
below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
Purity:
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained.
Amino-Acid Sequence :
The sequence of the first five N-terminal amino acids was determined and was found
to be Ala-Phe-Ser-Gly-Ser.
Dimers and aggregates:
Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity:
The bioactivity was measured by HMEC cell line, PEX can inhibit the transmembrane activity of HMEC under the stimulation of VEGF.
Endotoxin:
Less than 0.1 ng/ug (IEU/ug) of PEX.
Protein content:
Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm .
2. Analysis by RP-HPLC, using a calibrated solution of PEX as a Reference Standard.
Usage:
This material is offered by ProSpec-TechnoGene for research,
laboratory or further evaluation purposes.
Latest Publications:
1. Structural characterization and binding properties of the hemopexin-like domain of the matrixmetalloproteinase-19.
Protein Expr Purif 2005 Sep 21;
2. CD44 binding through the hemopexin-like domain is critical for its shedding by membrane-type 1 matrix metalloproteinase.
Oncogene 2005 Jan 27;24(5):859-68
3. Kinetic analysis of the binding of hemopexin-like domain of gelatinase B cloned and expressed in Pichia pastoris to tissue inhibitor of metalloproteinases-1.
J Protein Chem 2003 Aug;22(6):509-14
4. CD44 directs membrane-type 1 matrix metalloproteinase to lamellipodia by associating with its hemopexin-like domain.
EMBO J 2002 Aug 1;21(15):3949-59
5. Modulation of the catalytic activity of neutrophil collagenase MMP-8 on bovine collagen I. Role of the activation cleavage and of the hemopexin-like domain.
J Biol Chem 2002 Jun 28;277(26):23123-30
6. Specific, high affinity binding of tissue inhibitor of metalloproteinases-4 (TIMP-4) to the COOH-terminal hemopexin-like domain of human gelatinase A. TIMP-4 binds progelatinase A and the COOH-terminal domain in a similar manner to TIMP-2.
J Biol Chem 1997 Jun 13;272(24):15496-500