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Merck - PF038-10UG - MMP-9, Proenzyme, Human, Recombinant, CHO Cells

  • MFR SKU: PF038-10UG
  • ITEM #: 6280186
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ITEM #: 6280186
Proenzyme MMP-9^ Substrate Specificity: Collagen (Type IV, V, VII, X), elastin, gelatin (Type I, V). Form: Liquid. Species Reactivity: Human. Applications: Substrate Cleavage Assay, Western Blot, Zymography. Comments: Simple activation step is included in protocol.
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Merck - PF038-10UG - MMP-9, Proenzyme, Human, Recombinant, CHO Cells

  • Item #: 6280186
  • MFR SKU: PF038-10UG
Technical Specs
  • Overview Recombinant, human pro-MMP-9 expressed in CHO cells. A portion (less than 10%) of the enzyme may be seen as a naturally-occurring dimer. Dimerization does not interfere with activation. May contain up to 10% TIMP proteins. During storage, a small portion (less than 10%) of the enzyme may also become activated. Requires activation prior to use. A simple activation protocol is included.
  • Catalogue Number PF038
  • Brand Family Calbiochem®
  • Synonyms Gelatinase B, 92 kDa Gelatinase, Matrix Metalloproteinase 9, Type IV Collagenase
  • References Parsons, S.L., et al. 1997. Br. J. Surg. 84 , 160.
  • Backstrom, J.R., et al. 1996. J. Neuro. 16 , 7910.
  • Lim, G.P., et al. 1996. J Neurochem. 67 , 251.
  • Sang, Q.X., et al. 1995. Biochim. Biophys. Acta. 1251 , 99.
  • Kenagy, R.D. and Clowes, A.W. 1994. in Inhibition of Matrix Metalloproteinases: Therapeutic Potential. Greenwald, R.A. and Golub L.M., Eds.: 462-465.
  • Zempo, N., et al. 1994. J. Vasc. Surg. 20 , 209.
  • Birkedal-Hansen, H. 1993. J. Periodontol. 64 , 474.
  • Stetler-Stevenson, W.G., et al. 1993. FASEB J. 7 , 1434.
  • Delaisse, J-M. and Vaes, G. 1992. in Biology and Physiology of the Osteoclast. B.R. Rifkin & C.V. Gay, Eds.: 290-314.
  • Jeffrey, J.J. 1992. in Wound Healing: Biochemical and Clinical Aspects. R.F. Diegelmann and W.J. Lindblad, Eds.: 177-194.
  • Jeffrey, J.J. 1991. Semin. Perinatol. 15 , 118.
  • Liotta, L.A., et al. 1991. Cell 64 , 327.
  • Harris, E. 1990. N. Engl. J. Med. 322 , 1277.
  • Activity >1, 300 pmoles/min/µg
  • Unit of Definition Specific activity is determined using 10 μ M (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3-[2, 4-dinitrophenyl]-L-2, 3-diaminopropionyl)-Ala-Arg-NH (excitation 320 nm, emission 405 nm), and 20 ng enzyme in 100 μl of 50 mM Tris-HCl, pH 7.5, 10 mM CaCl, 150 mM NaCl, and 0.05% BRIJ<sup>®</sup>-35 Detergent at room temperature.
  • EC number 3.4.24.35
  • Form Liquid
  • Formulation In 150 mM NaCl, 50 mM Tris-HCl, 10 mM CaCl, 0.05% BRIJ ® -35 Detergent, pH 7.5.
  • Preservative None
  • Quality Level MQ100
  • Application References Zymography Xia, T., et al. 1996. Biochim. Biophys. Acta. 1293 , 259. Kleiner, D.E. and Stetler-Stevenson W.G. 1994. Anal. Biochem. 218 , 325. Heussen, C. and Dowdle, E.B. 1980. Anal. Biochem. 102 , 196. Substrate Cleavage Assay Xia, T., et al. 1996. Biochim. Biophys. Acta. 1293 , 259.
  • Key Applications Immunoblotting (Western Blotting)
  • Substrate Cleavage Assay
  • Zymography
  • Application Notes Immunoblotting (1 µg protein/lane)
  • Substrate Cleavage Assay (1 µg protein/lane, see application references)
  • Zymography (1 µg protein/lane, see application references)
  • Application Comments MMP-9 Proenzyme can be measured by its ability to degrade gelatin in a zymogram. 0.5 ng of enzyme is sufficient to visualize degraded gelatin with coomassie blue stain. The specific activity as measured with 10 µ M (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3-[2, 4-dinitrophenyl]-L-2, 3-diaminopropionyl)-Ala-Arg-NH 2 (excitation 320 nm, emission 405 nm), and 20 ng enzyme in 100 µl of 50 mM Tris-HCl, pH 7.5, 10 mM CaCl 2 , 150 mM NaCl, and 0.05% Brij-35 at room temperature, is >1, 300 pmoles/min/µg. To activate MMP-9 proenzyme, prepare p-aminophenylmercuric acetate (APMA) concentrate in dimethylsulfoxide (DMSO). Add APMA to MMP-9 proenzyme to give a final APMA concentration of 1 mM. Incubate at 37° C for 16 to 24 h.
  • Purity >90% by SDS-PAGE
  • Concentration Label Please refer to vial label for lot-specific concentration
  • Ship Code Dry Ice Only
  • Toxicity Standard Handling
  • Storage ≤ -70° C
  • Avoid freeze/thaw Avoid freeze/thaw
  • Do not freeze Ok to freeze
  • Special Instructions Following initial thaw, aliquot and freeze (-70° C).
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