Recombinant Human Monoglyceride Lipase Protein, CF

Catalog # Availability Size / Price Qty
7930-MG-010
Product Details
Citations (1)
FAQs
Reviews

Recombinant Human Monoglyceride Lipase Protein, CF Summary

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze 4-Nitrophenyl butyrate. The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Monoglyceride Lipase protein
Pro2-Pro303, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
34 kDa
SDS-PAGE
34-37 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rhMGLL to 0.8 ng/μL in Assay Buffer.
  2. Dilute Substrate to 2 mM using 40% ethanol.
  3. 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.
  4. Read at an absorbance of 410 nm in kinetic mode for 5 minutes.
  5. 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).

Per Well
  • rhMGLL: 0.04 μg
  • PNPB: 1 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Karlsson, M. et al. 1997, J. Biol. Chem. 272:27218.
  2. Nomura, D. K. et al. 2011, Science 334:809.
  3. Pan, B. et al. 2011, J. Neurosci. 31:13420.
  4. Chen, R. et al. 2012, Cell Rep. 2:1329.
Entrez Gene IDs
11343 (Human); 23945 (Mouse); 29254 (Rat)
Alternate Names
HUK5; HU-K5EC 3.1.1.23; Lysophospholipase homolog; Lysophospholipase-like; MAGL; MGLHUK5; MGLL; Monoacylglycerol lipase; Monoglyceride Lipase

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

  1. Free-energy studies reveal a possible mechanism for oxidation-dependent inhibition of MGL
    Sci Rep, 2016;6(0):31046.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay

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