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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
Ordering Information
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
Ordering Information
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
Ordering Information
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
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-103-C010   10 µg 560.00 USD Add To Cart
Product Specification
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli.
CONCENTRATION: 0.2mg/ml
PURITY: ≥95% (SDS-PAGE)
FORMULATION: Liquid. In 50mM TRIS-HCl, pH 7.5  containing 150mM NaCl, 5mM CaCl2, 1% glycerol and 0.1% Triton X-100.
SPECIFIC ACTIVITY: ≥75mU/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
LONG TERM STORAGE: -80°C
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-16 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) = (ΔFMca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg x  CMca-Pro-Leu x Vtotal) / (1000 x FMca-Pro-Leu x Venzyme)
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-16 is inhibited by tissue inhibitors of MMP-2 (TIMP-2) and by chelators of divalent cations like EDTA or o-phenanthroline.

Swiss-Prot link P51512: MMP-16 (human) (precursor)
AfCS Siganlling Gateway link A001468: MMP-16 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-104 Revised 04-Mar-08
MMP-17 (Catalytic Domain) (human) (recombinant) (His)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-104-C010   10 µg 560.00 USD Add To Cart
Product Specification
EC: 3.4.24.-
SOURCE/HOST: Produced in E. coli. Fused at the C-terminus to a His-tag.
CONCENTRATION: 0.2mg/ml
PURITY: ≥90% (SDS-PAGE)
FORMULATION: Liquid. 50mM TRIS-HCl, pH 7.5 containing 150mM NaCl, 5mM CaCl2, 1% glycerol and 0.1% Triton X-100.
SPECIFIC ACTIVITY: ≥6mU/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
LONG TERM STORAGE: -80°C
Product Description
The catalytic domain of human MMP-17 is produced by tryptic activation of a recombinant soluble proform.
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-17 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 (0.016-0.08μM). 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 8 to 10µl of the activation mixture are than added and the increase in fluorescence is recorded over a time interval between 2 and 15 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-17 is inhibited by tissue inhibitors of MMP-1 (TIMP-1) and by chelators of divalent cations like EDTA.


Swiss-Prot link Q9ULZ9: MMP-17 (human) (precursor)
AfCS Signalling Gateway link A001469: MMP-17 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-201-105 Revised 08-Jul-08
MMP-24 (Catalytic Domain) (human) (recombinant) (His)
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PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY MMPs
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-105-C010   10 µg 560.00 USD Add To Cart
Product Specification
MW: ~23kDa
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 and 5mM CaCl2.
SPECIFIC ACTIVITY: ≥120mU/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
LONG TERM STORAGE: -80°C
Product Description
Activation generates two molecular species. One species begins at the N-terminus with Tyr156 of full-length MMP-24, while the other species is two aa residues shorter and starts with Leu158. Both species extend to Ser351 of MMP-24 and contain an additional His-tag sequence at the C-terminus.
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-24 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-24 is inhibited by tissue inhibitors of MMP-2 (TIMP-2) and by chelators of divalent cations like EDTA or o-phenanthroline.

Swiss-Prot link Q9Y5R2: MMP-24 (human) (precursor)
AfCS Signalling Gateway link A001474: MMP-24 (mouse)
Further Categories Containing This Product:
Enzymes • Recombinant Proteins / Fusion Proteins
 
 
ALX-430-094 Revised 04-Jul-07
Monastrol
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SYNONYMS 4-(3-Hydroxyphenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydro-4H-pyrimidin-5-carboxylic acid ethyl ester
PRODUCT LINE Cell Cycle
PRODUCT CATEGORY Cell Cycle Blockers & Inhibitors / Related Products
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-430-094-M001   1 mg 60.00 USD Add To Cart
ALX-430-094-M005   5 mg 180.00 USD Add To Cart
Product Specification
FORMULA: C14H16N2O3S
MW: 292.4
CAS NUMBER: 329689-23-8
PURITY: ≥95%
APPEARANCE: White to light yellow solid.
SOLUBILITY: Soluble in DMSO or 100% ethanol.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.

Product Description
Cell permeable non-tubulin-interacting mitosis inhibitor. Blocks mitosis (IC50=14µM) by binding to the mitotic kinesin Eg5.
Product Specific Literature References
Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen: T.U. Mayer, et al.; Science 286, 971 (1999) Abstract
A combinatorial approach to recognition of chirality: preparation of highly enantioselective aryl-dihydropyrimidine selectors for chiral HPLC: K. Lewandowski, et al.; J. Comb. Chem. 1, 105 (1999) Abstract
Evidence that monastrol is an allosteric inhibitor of the mitotic kinesin Eg5: Z. Maliga, et al.; Chem. Biol. 9, 989 (2002) Abstract
Inhibition of a mitotic motor protein: where, how, and conformational consequences: Y. Yan, et al.; J. Mol. Biol. 335, 547 (2004) Abstract
Monastrol, a selective inhibitor of the mitotic kinesin Eg5, induces a distinctive growth profile of dendrites and axons in primary cortical neuron cultures: S.Y. Yoon, et al.; Cell Motil. Cytoskeleton 60, 181 (2005) Abstract
Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein: T. Peters, et al.; Naunyn Schmiedebergs Arch. Pharmacol. 372, 291 (2006) Abstract
A pathway of structural changes produced by monastrol binding to Eg5: Z. Maliga, et al.; J. Biol. Chem. 281, 7977 (2006) Abstract
Allosteric inhibition of kinesin-5 modulates its processive directional motility: B.H. Kwok, et al.; Nat. Chem. Biol. 2, 480 (2006) Abstract
Further Categories Containing This Product:
Kinesin & Kinesin-like Proteins / Related Products
 
 
ALX-803-311 Revised 17-Mar-08
Monoclonal Antibody to Aggrecan (5C5)
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SYNONYMS anti-Chondroitin Sulfate Proteoglycan MAb (5C5)
PRODUCT LINE Cytoskeleton
PRODUCT CATEGORY Proteoglycans / Related Products
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-803-311-R100   100 µl 310.00 USD Add To Cart
Product Specification
SPECIES CROSSREACTIVITY:
Human
Others
CLONE: 5C5
ISOTYPE: Mouse IgG1
FORMULATION: