Recombinant Human PLA2G4A Protein, CF

R&D Systems | Catalog # 6659-PL

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human PLA2G4A Protein (6659-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_077734

Applications

Enzyme Activity
View all PLA2G4A Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human PLA2G4A protein
Met1-Ala749, with a C-terminal 6-His tag

Purity

>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Ser2

Predicted Molecular Mass

86 kDa

SDS-PAGE

85-95 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 1-Hexadecanoyl-2-(1-pyrene-decanoyl)-sn-glycero-3-phosphocholine.
The specific activity is >15 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6659-PL
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, TCEP,Brij-35 and Glycerol.
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: PLA2G4A

Cytosolic Phospholipase A2a (PLA2G4A) belongs to the group IV phospholipase A2 family (1). These enzymes hydrolyze the ester bond at the second position (sn-2) of membrane glycerophospholipids. The group IV phospholipase A2 (PLA2G4) family is comprised of six intracellular enzymes (2). PLA2G4A has a preference for arachidonic acid in the sn-2 position of phospholipids indicating its involvement in the metabolism of eicosanoids. Its active site is characterized by a serine and aspartic acid dyad. The overall structural feature displays two domains, a lipid binding C2 domain and a catalytic a/b hydrolase domain. PLA2G4A translocation to the membrane and activation is facilitated by a calcium‑ion dependent conformational change in its C2 domain. In addition, phosphorylation of serine residues in PLA2G4A by protein kinases may be another regulatory mechanism for promoting the release of arachidonic acid. The knockout mouse studies of this enzyme indicate that the enzyme may play a major role in inflammatory diseases. Genetic ablation or reduction of PLA2G4A protected human amyloid precursor protein (APP) related pathogenesis from a mouse model (3). Mouse model studies of brain and lung cancer indicates that PLA2G4A may play a key regulatory role in angiogenesis of these tumors (4).

References

  1. Burke, J. E. and E.A. Dennis (2009) J. Lipid Res. 50:S237.
  2. Ghosh, M. et al. (2006) Prog. Lipid Res. 45:487.
  3. Sanchez-Mejia, R.O. et al. (2008) Nat. Neuroscience 11:1311.
  4. Linkous, A.G. et al. (2010) J. Natl. Cancer Inst. 102:1398.

Long Name

Phospholipase A2 Group IV A

Alternate Names

cPLA2, cPLA2-alpha

Entrez Gene IDs

5321 (Human); 18783 (Mouse); 24653 (Rat)

Gene Symbol

PLA2G4A

UniProt

NP_077734

Additional PLA2G4A Products

Product Documents for Recombinant Human PLA2G4A Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human PLA2G4A Protein, CF

For research use only

Citations for Recombinant Human PLA2G4A Protein, CF

Customer Reviews for Recombinant Human PLA2G4A Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human PLA2G4A Protein, CF and earn rewards!

Have you used Recombinant Human PLA2G4A Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human PLA2G4A Protein, CF (6659-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 Human PLA2G4A (rhPLA2G4A) (Catalog # 6659-PL)
  • Substrate: 1-Hexadecanoyl-2-(1-Pyrenedecanoyl)-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 rhPLA2G4A to 2 µg/mL in Assay Buffer.
  3. Dilute Substrate to 10 µM in Substrate Buffer.
  4. Load 50 µL of 2 µg/mL rhPLA2G4A into a black well plate, and start the reaction by adding 50 µL of 10 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 10 µM 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:
  • rhPLA2G4A: 0.10 µg
  • Substrate: 5 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes