Merck
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M9195

Sigma-Aldrich

Matrix Metalloproteinase-1 human

recombinant, expressed in NSO cells, >95% (SDS-PAGE), buffered aqueous solution

Synonym(s):
Collagenase-1, Interstitial Collagenase, MMP-1
Enzyme Commission number:
Número MDL:
NACRES:
NA.32

recombinante

expressed in NSO cells

Nível de qualidade

200

teor

>95% (SDS-PAGE)

forma

buffered aqueous solution

enzyme activity

≥400 pmol/min-μg

peso molecular

apparent mol wt 52-55 kDa

nº de adesão UniProt

enviado em

dry ice

temperatura de armazenamento

−70°C

Gene Information

human ... MMP1(4312)

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Ações bioquímicas/fisiológicas

MMP-1 initiates the breakdown of interstitial collagens.

forma física

Supplied as a 0.2 μm filtered solution of 25 mM MES, 10 mM calcium chloride, 150 mM sodium chloride, 0.05% Brij® L23, pH 5.5.

Nota de análise

The biological activity is measured by its ability to cleave a fluorogenic peptide substrate.

Outras notas

Activity Assay Protocol

Materials:
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
  • Recombinant Human MMP-1 (rhMMP-1) (Cat. No. M9195)
  • p-aminophenylmercuric acetate (APMA), (Cat. No. A9563), 100 mM stock in DMSO
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2
  • F16 Black Maxisorp Plate
  • Fluorescent Plate Reader or equivalent

Assay:
1. Dilute rhMMP-1 to 50 μg/mL in Assay Buffer.
2. Activate 50 μg/mL rhMMP-1 by adding APMA to a final concentration of 1 mM.
3. Incubate at 37 °C for 2 hours.
4. Dilute activated rhMMP-1 to 1 ng/μL in Assay Buffer.
5. Dilute Substrate to 20 μM in Assay Buffer.
6. Load into a black well plate 50 μL of 1 ng/μL rhMMP-1 and start the reaction by adding 50 μL of 20 μM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer, 50 μL Substrate, and no rhMMP-1.
7. Read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
8. Calculate specific activity:
     Specific Activity (pmol/min/μg) =Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) / amount of enzyme (μg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH.

Final Assay Conditions:

Per Well:
rhMMP-1: 0.050 μg
Substrate: 10 μM

Informações legais

Brij is a registered trademark of Croda International PLC

Código de classe de armazenamento

12 - Non Combustible Liquids

WGK

WGK 2

Ponto de fulgor (ºF)

Not applicable

Ponto de fulgor (ºC)

Not applicable

Equipamento de proteção individual

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

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