Recombinant Mouse PLA2G2A Protein, CF

R&D Systems | Catalog # 4925-PL

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse PLA2G2A Protein (4925-PL)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

NP_001076000

Applications

Enzyme Activity
View all PLA2G2A Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse PLA2G2A protein
Asn22-Cys146, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Asn22

Predicted Molecular Mass

15 kDa

SDS-PAGE

17 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 1-Hexadecanoyl-2-(1-pyrene-decanoyl)-sn-glycero-3-phosphocholine.
The specific activity is >50 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

4925-PL
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: PLA2G2A

Secretory Phospholipase A2 is an enzyme that hydrolyses the sn-2 ester bond of phospholipids, generating non-esterified free fatty acids and lysophospholipids (1‑3). PLA2G2A is a calcium-dependent phospholipase expressed in many cell types participating in inflammation-associated cellular responses, including platelets, neutrophils, and mast cells. It may function as an enzymatic component of the host defense mechanism. For example, human tears contain a high concentration of PLA2G2A, a principal bactericidal factor against Gram-positive bacteria in this fluid. It may play a role in cell proliferation through binding a receptor on the cell membrane. PLA2G2A has been shown to have pro-atherogenic properties both in the circulation and within the arterial wall (4). It is an acute phase protein expressed in response to a variety of pro-inflammatory cytokines. Circulating levels of sPLA2G2A are higher in coronary artery disease (CAD) patients and are associated with increased risk of future CAD (5).

References

  1. Webb, N. R. (2005) Cur. Opin. Lipid. 16:341.
  2. Triggiani, M. et al. (2005) J. Allergy Clin. Immunol. 116:1000.
  3. Murakami, M. and Kudo, I. (2004) Biol. Pharm. Bull. 27:1158.
  4. de Beer, F. C. and Webb, N. R. (2006) Arterioscler. Thromb. Vasc. Biol. 26:1421.
  5. Wootton, P. T. E. et al. (2006) Human Mol. Genet. 15:355.

Long Name

Phospholipase A2 Group IIA

Alternate Names

Mom1, PLA2B, PLA2L, RASF-A

Entrez Gene IDs

5320 (Human); 18780 (Mouse); 29692 (Rat)

Gene Symbol

PLA2G2A

UniProt

NP_001076000

Additional PLA2G2A Products

Product Documents for Recombinant Mouse PLA2G2A Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse PLA2G2A Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse PLA2G2A Protein, CF (4925-PL):

Materials
  • Assay Buffer: 50 mM Tris, 500 mM NaCl, 1 mg/mL BSA, pH 8.5.
  • Substrate Buffer: 50 mM Tris, 500 mM NaCl, 20 mM CaCl2, 1 mg/mL BSA, pH 8.5.
  • Recombinant Mouse Phospholipase A2 Group IIA/PLA2G2A (rmPLA2G2A) (Catalog # 4925-PL)
  • Substrate: 1-hexadecanoyl-2-(1-pyrene-decanoyl)-sn-glycero-3-phosphocholine (Invitrogen, Catalog # H361), Dilute to 2 mM in DMSO, then dilute to a final stock concentration of 400 µM in ethanol
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Thaw Substrate at 37 °C for five minutes. Mix well.
  2. Dilute rmPLA2G2A to 2 ng/µL in Assay Buffer.
  3. Dilute substrate PGPC to 20 µM in Substrate Buffer.
  4. Load in plate 50 µL of 2 ng/µL rmPLA2G2A, and start the reaction by adding 50 µL of 20 µM Substrate to each well. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
  5. Read at excitation and emission wavelengths of 345 nm and 395 nm (top read), respectively, in kinetic mode for 5 minutes.
  6. 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 1-pyrenedecanoic acid (Invitrogen, Catalog # P31).

Per Well:
  • rmPLA2G2A: 0.1 µg
  • Substrate: 10 µM

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