Recombinant Human Monoglyceride Lipase Protein, CF
Recombinant Human Monoglyceride Lipase Protein, CF Summary
Product Specifications
Pro2-Pro303, with an N-terminal Met and 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
7930-MG
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij-35 and EDTA. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM Tris, pH 8.0
- Ethanol, Absolute
- Recombinant Human Monoglyceride Lipase (rhMGLL) (Catalog # 7930-MG)
- Substrate: 4-Nitrophenyl butyrate (PNPB) (Sigma, Catalog # N9876), 100 mM stock in Acetone
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhMGLL to 0.8 ng/μL in Assay Buffer.
- Dilute Substrate to 2 mM using 40% ethanol.
- Load 50 μL of 0.8 ng/μL rhMGLL into the plate, and start the reaction by adding 50 μL of 2 mM Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 2 mM Substrate.
- Read at an absorbance of 410 nm in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 4-Nitrophenol (4-NP) (Sigma, Catalog # 241326).
- rhMGLL: 0.04 μg
- PNPB: 1 mM
Reconstitution Calculator
Background: Monoglyceride Lipase
Monoacylglycerol lipase, also known as MAG lipase and MGLL, is a member of the serine hydrolase superfamily with the typical serine hydrolase catalytic triad (1). It converts monoacylglycerides to free fatty acids and glycerol. An important physiological process performed by MGLL is the breakdown of the endocannabinoid 2‑arachidonoylglycerol (2-AG) by hydrolysis to provide the arachidonic acid precursor for pro‑inflammatory eicosanoid synthesis (2). MGLL knock‑out mice exhibited enhanced learning and improved performance in novel object recognition due the increased level of 2‑AG (3). In Alzheimer’s disease, inactivation of MGLL suppressed accumulation of b‑amyloid and reduced the expression of b‑site amyloid precursor protein cleaving enzyme 1 (BACE1) (4).
- Karlsson, M. et al. 1997, J. Biol. Chem. 272:27218.
- Nomura, D. K. et al. 2011, Science 334:809.
- Pan, B. et al. 2011, J. Neurosci. 31:13420.
- Chen, R. et al. 2012, Cell Rep. 2:1329.
Product Specific Notices
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.Citation for Recombinant Human Monoglyceride Lipase Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
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Free-energy studies reveal a possible mechanism for oxidation-dependent inhibition of MGL
Sci Rep, 2016-08-08;6(0):31046.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay
FAQs
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