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ALX-200-424 Revised 04-Dec-04
MMP-9/Lipocalin Complex (human)
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SYNONYMS Matrix Metalloproteinase 9/Lipocalin Complex (human)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-200-424-C005   5 µg 166.00 USD Add To Cart
Product Specification
MW: 130kDa.
EC: 3.4.24.35 (MMP-9)
SOURCE/HOST: Isolated from stimulated human neutrophil granulocytes (buffy coat). Disulfide-bridged complex of the monomer and a 25kDa protein of the lipocalin family. Requires activation.
PURITY: ≥95% (SDS-PAGE, Western blot)
FORMULATION: Liquid. 5µg protein in 200mM NaCl, 50mM Tris-HCl pH 7.0, 5mM CaCl2, 1µM ZnCl2, 0.05% BRIJ35, and 0.05% sodium azide.
SPECIFIC ACTIVITY: ≥1'500mU/mg protein. One unit is defined as the amount of enzyme that hydrolyzes 1µmol of Dnp-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min. at 37°C, pH 7.0 (Y. Masui, et al.; Biochem. Med. 17, 215 (1977)).
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles.
Product Specific Literature References
Synthetic substrates for vertebrate collagenase: Y. Masui, et al.; Biochem. Med. 17, 215 (1977) Abstract
A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
Progelatinase B forms from human neutrophils. complex formation of monomer/lipocalin with TIMP-1: H. Kolkenbrock, et al.; Biol. Chem. 377, 529 (1996) Abstract
Human neutrophil gelatinase and associated lipocalin in adult and localized juvenile periodontitis: U. Westerlund, et al.; J. Dent. Res. 75, 1553 (1996) Abstract
General Information
MMP-9 hydrolyzes the extracellular matrix, e.g. collagen types IV, V and IX and gelatin. Product 200-424 represents the 130kDa disulfide-bridged complex of the monomer and a 25kDa protein of the lipocalin family and displays homology with the α2-macroglobulin-related protein from rats. The human MMP-9/lipocalin complex was shown to be associated with adult and localized juvenile periodontitis.
BACKGROUND/TECHNICAL INFORMATION Swiss-Prot link: MMP-9 (human) (precursor) (P1480), Lipocalin (human) (precursor) (P80188)
Further Categories Containing This Product:
Natural Proteins • Enzymes
 
 
ALX-200-425 Revised 04-Mar-05
MMP-9/Lipocalin/TIMP-1 Complex (human)
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SYNONYMS Matrix Metalloproteinase 9/Lipocalin/Tissue Inhibitor of Metalloproteinase 1 Complex (human)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-200-425-C005   5 µg 166.00 USD Add To Cart
Product Specification
EC: 3.4.24.35 (MMP-9)
SOURCE/HOST: Isolated from stimulated human neutrophil granulocytes (buffy coat). Requires activation.
PURITY: ≥95% (SDS-PAGE, Western blot)
FORMULATION: Liquid. 5µg protein in 200mM NaCl, 50mM Tris-HCl pH 7.0, 5mM CaCl2, 1µM ZnCl2, 0.05% BRIJ35 and 0.05% sodium azide.
SPECIFIC ACTIVITY: ≥400mU/mg protein. One unit is defined as the amount of enzyme that hydrolyzes 1µmol Dnp-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH per min. at 37°C, pH 7.0 (Y. Masui, et al.; Biochem. Med. 17, 215 (1977)).
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles.
Product Description
This product represents the ternary complex of the MMP-9 monomer, lipocalin and TIMP-1 and behaves like the progelatinase B/TIMP-1 complex, that inhibits active MMP's and, after activation acts as a gelatinase with reduced activity.
Product Specific Literature References
Synthetic substrates for vertebrate collagenase: Y. Masui, et al.; Biochem. Med. 17, 215 (1977) Abstract
A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
Progelatinase B forms from human neutrophils. complex formation of monomer/lipocalin with TIMP-1: H. Kolkenbrock, et al.; Biol. Chem. 377, 529 (1996) Abstract
General Information
MMP-9 hydrolyzes the extracellular matrix, e.g. collagen types IV, V and IX and gelatin.
BACKGROUND/TECHNICAL INFORMATION Swiss-Prot link: MMP-9 (human) (precursor) (P14780), Lipocalin (human) (precursor) (P80188), TIMP-1 (human) (precursor) (P01033)
Further Categories Containing This Product:
Tissue Inhibitors of Matrix Metalloproteinases [TIMPs] / Related Products • Natural Proteins • Enzymes
 
 
ALX-201-097 Revised 04-Sep-08
MMP-13 (human) (recombinant) (His)
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SYNONYMS Procollagenase-3 (human) (recombinant) (His)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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ALX-201-097-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~52kDa (unglycosylated)
EC: 3.4.24.-
SOURCE/HOST: Produced in Sf9 cells.
CONCENTRATION: 0.2mg/ml
PURITY: ≥95% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2, 0.05% Brij-35.
SPECIFIC ACTIVITY: ≥200mU/mg protein. One unit is defined as the amount of enzyme that hydrolyzes 1µmol Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 per min. at 38°C, pH 7.5.
APPLICATION: Antigen standard in immunochemical analyses. The active enzyme may be used to study the degradation of extracellular matrix proteins, to screen inhibitors of matrix metalloproteinases and to characterize inhibitor actions.
SHIPPING: SHIPPED ON DRY ICE
SHORT TERM STORAGE: -20°C
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles.
General Information
MMP-13 is inhibited by tissue inhibitors of matrix metalloproteinases (TIMP) and by chelators of divalent cations as EDTA or o-phenanthroline.
BACKGROUND/TECHNICAL INFORMATION Activation of MMP-13 proenzyme and measurement of catalytic activity

1 Preparation and stability of solutions:
∙ APMA-solution: 40mM p-aminophenyl mercuric acetate (APMA) in dimethylsulfoxide. Store at -20°C.
∙ Peptide hydrolysis buffer: 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2, 0.025% Brij 35. Solution is stable for several weeks at 4°C.
∙ Stock solution of peptide substrate: 100μM solution of Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg in 20% dimethylsulfoxide. Store at -20°C.
∙ Stock solution of unquenched peptide: 10μM solution of (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-NH2 (Mca-Pro-Leu) in 20% dimethylsulfoxide. Store at -20°C.

2 Activation:
An aliquot of 19.5μl MMP-13 proenzyme is mixed with 0.5µl APMA solution and the mixture is incubated for 90 min. at 37°C.

3 Assay protocol:
The activity of MMP-13 is measured fluorimetrically with a synthetic internally quenched fluorescent substrate according to Knight et al. [1].
An excitation wavelength of 328nm and an emission wavelength of 393nm are set in an appropriate fluorimeter. The instrument is calibrated with the unquenched peptide Mca-Pro-Leu at a concentration corresponding to between 2 and 10% hydrolysis of the protease substrate. Kinetic reactions are conveniently carried out in a constant volume of 2.5ml. The substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg is diluted in peptide hydrolysis buffer to a concentration of  0.8µM and equilibrated at a temperature of 37°C. Aliquots of 1 to 2µl of the activation mixture are than added and the increase in fluorescence is recorded over a time interval between 2 and 12 min. Activity units per ml enzyme solution are calculated according to the following equation:

Activity (U/ml) = (CMca-Pro-Leu/FMca-Pro-Leu) x (ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg/Venzyme) x Vtotal

CMca-Pro-Leu: Concentration of Mca-Pro-Leu used for calibration of the fluorimeter (mmoles/ml)
FMca-Pro-Leu: Fluorescence of Mca-Pro-Leu at the concentration CMca-Pro-Leu used for fluorimeter calibration
ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg: Change in fluorescence during peptide hydrolysis per min
Vtotal: Volume of peptide hydrolysis reaction (2.5ml)
Venzyme: Volume of added enzyme (0.001 to 0.002ml)

Due to autoproteolytic activity minor bands of activated enzyme may be visible in the preparation.

[1] A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992)

Swiss-Prot link P45452: MMP-13 (human) (precursor)
AfCS Signalling Gateway link A001465: MMP-13 (mouse)
General Literature References
[1] A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-100 Revised 02-Oct-08
MMP-14 Proenzyme, Soluble (human) (recombinant) (His)
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SYNONYMS MT1-MMP Proenzyme, Soluble (human) (recombinant) (His)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-100-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~58kDa.
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli. Prodomain, catalytic domain and hemopexin domain of human MMP-14 proenzyme (aa 1-501) are fused to a His-tag (Thr-(His)6).
CONCENTRATION: 0.2mg/ml
PURITY: ≥80% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5, containing 150mM NaCl and 5mM CaCl2.
APPLICATION: Used as antigen for antibody generation and as antigen standard in immunoassays. The proenzyme can be activated with trace amounts of MMP-14 catalytic domain [1, 2]. Active MMP-14 is used to study the activation of progelatinase A (matrixmetalloproteinase 2) and the degradation of proteins of the extracellular matrix. The enzyme allows screening of matrixmetalloproteinase inhibitors and characterization of inhibitor action.
SHIPPING: SHIPPED ON DRY ICE
SHORT TERM STORAGE: -20°C
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable for several weeks when stored at -20°C.
HANDLING: Avoid freeze/thaw cycles.
Product Description
Active soluble MMP-14 is used to study the activation of MMP-2 (progelatinase A) and the degradations of proteins of the extracellular matrix, including fibrillar collagens. Proenzyme is activated by trace amounts of MMP-14 catalytic domain [1,2].
Product Specific Literature References
[1] Membrane-type matrix metalloproteinases 1 and 2 exhibit broad-spectrum proteolytic capacities comparable to many matrix metalloproteinases: M.P. d’Ortho, et al.; Eur. J. Biochem. 250, 751 (1997) Abstract
[2] The TIMP2 membrane type 1 metalloproteinase "receptor" regulates the concentration and efficient activation of progelatinase A. A kinetic study: G.S. Butler, et al.; J. Biol. Chem. 273, 871 (1998) Abstract; Full Text
General Information
MMP-14 is expressed in adult lung, placenta, kidney, ovaries, intestine, prostate and spleen. Increased amounts of the enzyme are found in tumor tissues as lung carcinoma, gastric carcinoma, colon, breast, head and neck carcinoma. It activates pro-MMP-2 and pro-MMP-13 by proteolytic cleavage of their prodomains. The ability to activate other MMPs provides potential for enhanced pericellular proteolysis in physiological and pathological processes. In particular, activation of pro-MMP-2 by MMP-14 is considered to contribute to local degradation of extracellular matrix during cell migration and proliferation. MMP-14 hydrolyzes also fibrillar collagens I, II and III into characteristic 3/4 and 1/4 fragments and it cleaves a number of other proteins of the extracellular matrix, among them fibronectin, vitronectin, laminin-1 and dermatan sulfate proteoglycan. The activity of MMP-14 is poorly inhibited by tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), but efficiently inhibited by TIMP-2 and TIMP-3. MMP-14 proenzyme is activated by removal of its prodomain. The reaction is catalyzed by furin, a subtilisin-type serine protease, which recognizes a motif of four basic aa located between prodomain and catalytic domain.
BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P50281: MMP-14 (human) (precursor)
AfCS Signalling Gateway link A001466: MMP-14 (mouse)
General Literature References
The TIMP2 membrane type 1 metalloproteinase "receptor" regulates the concentration and efficient activation of progelatinase A. A kinetic study: G.S. Butler, et al.; J. Biol. Chem. 273, 871 (1998) Abstract; Full Text
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-099 Revised 08-Jan-08
MMP-14 Prodomain (Catalytic Domain) (human) (recombinant) (His)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-099-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~31kDa
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli. Prodomain (catalytic domain) of human MMP-14 proenzyme (aa 1-265) is fused to a His-tag (Leu-Val-Thr-(His)6).
CONCENTRATION: 0.2mg/ml
PURITY: ≥90% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5, containing 150mM NaCl and 5mMCaCl2.
SPECIFIC ACTIVITY: ≥60mU/mg protein after trypsin activation. One unit is defined as the amount of enzyme that hydrolyzes 1µmol Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 per min. at 37°C, pH 7.5.
SHIPPING: SHIPPED ON DRY ICE
SHORT TERM STORAGE: -20°C
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable for several weeks when stored at -20°C.
HANDLING: Avoid freeze/thaw cycles.
Product Description
Active catalytic domain MMP-14 is used to study the activation of MMP-2 (progelatinase A) and the degradation of proteins of the extracellular matrix, including fibrillar collagens. MMP-14 is inhibited by tissue inhibitors of MMP-2 and MMP-3 and by chelators of divalent cations as EDTA or o-phenanthroline.
Product Specific Literature References
[1] A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION Activation of MMP-14 proenzyme and measurement of catalytic activity

1 Preparation and stability of solutions:
∙ Activation buffer: 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2. Store at -20°C.
∙ Trypsin solution: 50µg TPCK-trypsin per ml activation buffer. Store in aliquots at -20°C.
∙ Aprotinin solution: 1mg aprotinin per ml activation buffer. Store at -20°C.
∙ Peptide hydrolysis buffer: 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2, 0.025% Brij 35. Store at +4°C for several weeks.
∙ Stock solution of peptide substrate: 100µM solution of Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 in 20% DMSO. Store at -20°C
∙ Stock solution of unquenched peptide: 10µM solution of Mca-Pro-Leu-NH2 in 20% DMSO. Store at -20°C.

2 Activation:
An aliquot of 5µg MMP-14 proenzyme is mixed with 1µl trypsin solution in 100µl avtivation buffer and incubated for 12 min. at 25°C. Thereafter trypsin is inhibited by addition of 1µl aprotinin solution.

3 Assay protocol:
The activity of MMP-14 is measured fluorimetrically with a synthetic internally quenched fluorescent substrate according to Knight et al. [1].
An excitation wavelength of 328nm and an emission wavelength of 393nm are set in an appropriate fluorimeter. The instrument is calibrated with the unquenched peptide Mca-Pro-Leu at a concentration corresponding to between 2 and 10% hydrolysis of the protease substrate. Kinetic reactions are conveniently carried out in a constant volume of 2.5ml. The substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg is diluted in peptide hydrolysis buffer to a concentration of  0.8µM and equilibrated at a temperature of 37°C. Aliquots of 4 to 8µl of the activation mixture are than added and the increase in fluorescence is recorded over a time interval between 2 and 12 minutes. Activity units per ml enzyme solution are calculated according to the following equation:

Activity (U/ml) = (CMca-Pro-Leu/FMca-Pro-Leu) x (ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg/Venzyme) x Vtotal

CMca-Pro-Leu: Concentration of Mca-Pro-Leu used for calibration of the fluorimeter (µmoles/ml).
FMca-Pro-Leu: Fluorescence of Mca-Pro-Leu at the concentration CMca-Pro-Leu used for fluorimeter calibration.
ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg: Change in fluorescence during peptide hydrolysis per min.
Vtotal: Volume of peptide hydrolysis reaction (2.5ml).
Venzyme: Volume of added enzyme (0.002 to 0.004ml).

Due to autoproteolytic activity minor bands of activated enzyme may be visible in the preparation.

Swiss-Prot link P50281: MMP-14 (human) (precursor)
AfCS Signalling Gateway link A001466: MMP-14 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-098 Revised 02-Oct-08
MMP-14 (Catalytic Domain) (human) (recombinant)
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SYNONYMS Matrix Metalloproteinase 14 (Catalytic Domain) (human) (recombinant)
MT1-MMP (Catalytic Domain) (human) (recombinant)
Membrane-type 1 Matrix Metalloproteinase (Catalytic Domain) (human) (recombinant)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-098-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~20kDa (SDS-PAGE).
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli. Mature human MMP-14 (aa 89-265).
CONCENTRATION: 0.2mg/ml
PURITY: ≥95% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2.
SPECIFIC ACTIVITY: ≥140mU/mg protein. One unit is defined as the amount of enzyme that hydrolyzes 1µmol Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 per min. at 37°C, pH 7.5.
APPLICATION: Used to study the activation of progelatinase A (matrix metalloproteinase 2) and the degradation of proteins of the extracellular matrix. The enzyme allows screening of matrix metalloproteinase inhibitors and characterization of inhibitor action.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
HANDLING: Avoid freeze/thaw cycles.
General Information
MMP-14 is expressed in adult lung, placenta, kidney, ovaries, intestine, prostate and spleen. Increased amounts of the enzyme are found in tumor tissues as lung carcinoma, gastric caqrcinoma, colon, breast, head and neck carcinoma. 
MMP-14 is activated by removal of its prodomain. The reaction is catalyzed by furin, a subtilisin-type serine protease, which recognizes a motif of four basic amino acid residues located between prodomain and catalytic domain.
MMP-14 activates pro-MMP-2 and pro-MMP-13 by proteolytic cleavage of their prodomains. The ability of MMP-14 to activate other MMPs provides potential for enhanced pericellular proteolysis in physiological and pathological processes. In particular, activation of pro-MMP-2 by MMP-14 is considered to contribute to local  degradation of extracellular matrix during cell migration and proliferation. MMP-14 hydrolyzes also fibrillar collagens I, II and III into characteristic 3/4 and 1/4 fragments and it cleaves a number of other proteins of the extracellular matrix, among them fibronectin, vitronectin, laminin-1 and dermatan sulfate proteoglycan. The activity of MMP-14 is poorly inhibited by tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), but efficiently inhibited by TIMP-2 and TIMP-3.
BACKGROUND/TECHNICAL INFORMATION
Measurement of MT1-MMP activity

Preparation and stability of solutions
Peptide hydrolysis buffer: 50mM Tris-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2, 0.025% Brij 35.
Solution is stable for several weeks at 4°C.
Stock solution of peptide substrate: 100mM solution of Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg in 20% dimethylsulfoxide. Store solution at -20°C.
Stock solution of unquenched peptide: 10mM solution of (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-NH2 (Mca-Pro-Leu) in 20% dimethylsulfoxide. Store solution at -20°C.

Assay protocol
The activity of MT1-MMP catalytic domain is measured fluorimetrically with a synthetic internally quenched fluorescent substrate according to Knight et al. [1].
An excitation wavelength of 328nm and an emission wavelength of 393nm are set in an appropriate fluorimeter. The instrument is calibrated with the unquenched peptide Mca-Pro-Leu at a concentration corresponding to between 2 and 10% hydrolysis of the protease substrate. Kinetic reactions are conveniently carried out in a constant volume of 2.5ml. The substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg is diluted in peptide hydrolysis buffer to a concentration of 0.8µM and equilibrated at a temperature of 37°C. Aliquots of 2 to 4µl of matrix metalloproteinase are than added and the increase in fluorescence is recorded over a time interval between 2 and 12 min. Activity units per ml enzyme solution are calculated according to the following equation:

Activity U/ml = (cMca-Pro-Leu/FMca-Pro-Leu) x (ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg/venzyme) x Vtotal

 cMca-Pro-Leu :  Concentration of Mca-Pro-Leu used for calibration of the fluorimeter (mmoles/ml)
 FMca-Pro-Leu :  Fluorescence of Mca-Pro-Leu at the concentration c Mca-Pro-Leu  used for fluorimeter calibration
 ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg: Change in fluorescence during peptide hydrolysis per min
 V :  Volume of peptide hydrolysis reaction (2.5 ml)
  v :  Volume of added enzyme (0.002 ml to 0.004 ml)

Sequence
Y89 A I Q G L K W Q H N E I T F C I Q N Y T P K V G E Y A T Y E A I R K A F R V W E S A T P L R F R E V P Y A Y I R E G H E K Q A D I M I F F A E G F H G D S T P F D G E G G F L A H A Y F P G P N I G G D T H F D S A E P W T V R N E D L N G N D I F L V A V H E L G H A L G L E H S S D P S A I M A P F Y Q W M D T E N F V L P D D D R R G I Q Q L Y G G E S G265

Inhibitors
The catalytic domain of MMP-14 is inhibited by tissue inhibitors of MMP-2 and -3 (TIMP-2 and TIMP-3) and by chelators of divalent cations like EDTA or o-phenanthroline.

Swiss-Prot link P50281: MMP-14 (human) (precursor)
AfCS Signalling Gateway link A001466: MMP-14 (mouse)
General Literature References
[1] A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-096 Revised 07-Dec-04
MMP-14 (Hemopexin Domain) (human) (recombinant) (His)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-096-C020   20 µg 560.00 USD Add To Cart
Product Specification
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli.
APPLICATION: Detection and monitoring of recombinant MMP-14 hemopexin domain and its binding to low molecular weight ligands and macromolecules.
SHIPPING: SHIPPED ON DRY ICE
LONG TERM STORAGE: -80°C
General Information
BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P50281: MMP-14 (human) (precursor)
AfCS Signalling Gateway link A001466: MMP-14 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-101 Revised 04-Mar-08
MMP-15 (Catalytic Domain) (human) (recombinant)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-101-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~20kDa
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli.
CONCENTRATION: 0.2mg/ml
PURITY: ≥90% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5, containing 150mM NaCl, 5mM CaCl2, 50mM imidazol and 0.05% Brji-35.
SPECIFIC ACTIVITY: ≥30mU/mg protein. One unit is defined as the amount of enzyme that hydrolyzes 1µmol Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 per min. at 37°C, pH 7.5.
SHIPPING: SHIPPED ON DRY ICE
SHORT TERM STORAGE: -20°C
LONG TERM STORAGE: -80°C
USE/STABILITY: Stable for several weeks when stored at -20°C.
HANDLING: Avoid freeze/thaw cycles.
Product Specific Literature References
A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases: C.G. Knight, et al.; FEBS Lett. 296, 263 (1992) Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION

Measurement of catalytic activity

1 Preparation and stability of solutions:
· Peptide hydrolysis buffer: 50mM TRIS-HCl, pH 7.5, 150mM NaCl, 5mM CaCl2, 0.025% Brij 35. Solution is stable for several weeks at 4°C.
· Stock solution of peptide substrate: 100µM solution of Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg in 20% DMSO. Store at -20°C.
· Stock solution of unquenched peptide: 10µM solution of (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-NH2 (Mca-Pro-Leu) in 20% DMSO. Store at -20°C.

2 Assay protocol:
The activity of MMP-15 is measured fluorimetrically with a synthetic internally quenched fluorescent substrate according to Knight et al.
An excitation wavelength of 328nm and an emission wavelength of 393nm are set in an appropriate fluorimeter. The instrument is calibrated with the unquenched peptide Mca-Pro-Leu at a concentration corresponding to between 2 and 10% hydrolysis of the protease substrate. Kinetic reactions are conveniently carried out in a constant volume of 2.5ml. The substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg is diluted in peptide hydrolysis buffer to a concentration of 0.8µM and equilibrated at a temperature of 37°C. Aliquots of 2 to 4µl of the activation mixture are than added and the increase in fluorescence is recorded over a time interval between 2 and 12 minutes. Activity units per ml enzyme solution are calculated according to the following equation:

Activity (U/ml) = (CMca-Pro-Leu/FMca-Pro-Leu) x (ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg/Venzyme) x Vtotal

CMca-Pro-Leu: Concentration of Mca-Pro-Leu used for calibration of the fluorimeter (µmoles/ml).
FMca-Pro-Leu: Fluorescence of Mca-Pro-Leu at the concentration CMca-Pro-Leu used for fluorimeter calibration.
ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg: Change in fluorescence during peptide hydrolysis per min.
Vtotal: Volume of peptide hydrolysis reaction (2.5ml).
Venzyme: Volume of added enzyme (0.002 to 0.004ml).

3 Inhibitors:
The catalytic domain of MMP-15 is inhibited by tissue inhibitors of MMP-2 and -3 (TIMP-2 and TIMP-3) and by chelators of divalent cations like EDTA or o-phenanthroline.

Swiss-Prot link P51511: MMP-15 (human) (precursor)
AfCS Signalling Gateway link A001467: MMP-15 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-103 Revised 04-Mar-08
MMP-16 (Catalytic Domain) (human) (recombinant)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-103-C010   10 µg 560.00 USD Add To Cart
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