Matrix Metalloproteinase-1 human
SIGMA/M9195 - recombinant, expressed in NSO cells, >95% (SDS-PAGE), buffered aqueous solution
Synonym: Collagenase-1; Interstitial Collagenase; MMP-1
MDL Number: MFCD00130830
Product Type: Chemical
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| assay | >95% (SDS-PAGE) |
| enzyme activity | ≥400 pmol/min-μg |
| form | buffered aqueous solution |
| mol wt | apparent mol wt 52-55 kDa |
| Quality Level | 200  |
| recombinant | expressed in NSO cells |
| shipped in | dry ice |
| storage temp. | −70°C |
| UniProt accession no. | P03956  |
| Analysis Note: | The biological activity is measured by its ability to cleave a fluorogenic peptide substrate. |
| Biochem/physiol Actions: | MMP-1 initiates the breakdown of interstitial collagens. |
| Legal Information: | Brij is a registered trademark of Croda International PLC |
| Other Notes: | 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-D • 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 |
| Physical form: | 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Purity | >95% (SDS-PAGE) |
| activity | enzyme activity: ≥400 pmol/min-μg |
| Storage Temp. | −70°C |
| Enzyme Commission (EC) Number | 3.4.24.7 ( BRENDA | IUBMB ) |
| UNSPSC | 12352202 |