半岛bd体育手机客户端 名称:
Matrix Metalloproteinase-1 human
MMP-1
Interstitial Collagenase
Collagenase-1
Matrix Metalloproteinase-1 人
半岛bd体育手机客户端 介绍:
半岛bd体育手机客户端 说明
生化/生理作用
MMP-1 initiates the breakdown of interstitial collagens.
外形
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.
分析说明
The biological activity is measured by its ability to cleave a fluorogenic peptide substrate.
其他说明
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 ℃ 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 V
max* (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
法律信息
Brij is a registered trademark of Croda International PLC
基本信息
酶学委员会(EC)编号 |
3.4.24.7 ( BRENDA | IUBMB ) |
MDL编号 |
MFCD00130830 |
NACRES |
NA.32 |
半岛bd体育手机客户端 性质
质量水平 |
200 |
重组 |
expressed in NSO cells |
测定 |
>95% (SDS-PAGE) |
形式 |
buffered aqueous solution |
enzyme activity |
≥400 pmol/min-μg |
分子量 |
apparent mol wt 52-55 kDa |
UniProt登记号 |
P03956 |
运输 |
dry ice |
储存温度 |
?70℃ |
Gene Information |
human ... MMP1(4312) |
安全信息
储存分类代码 |
12 - Non Combustible Liquids |
WGK |
WGK 2 |
闪点(F) |
Not applicable |
闪点(C) |
Not applicable |
个人防护装备 |
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US) |