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Recombinant Human PLA2G4A Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

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Recombinant Human PLA2G4A Protein, CF Summary

Details of Functionality
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.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human PLA2G4A protein
Met1-Ala749, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Ser2
Protein/Peptide Type
Recombinant Enzymes
Gene
PLA2G4A
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
86 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
85-95 kDa, reducing conditions

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, TCEP,Brij-35 and Glycerol.
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human PLA2G4A Protein, CF

  • calcium-dependent phospholipid-binding protein
  • cPLA2
  • cPLA2-alpha
  • lysophospholipase
  • MGC126350
  • phosphatidylcholine 2-acylhydrolase
  • Phospholipase A2 group IVA
  • phospholipase A2, group IVA (cytosolic, calcium-dependent)
  • PLA2G4A
  • PLA2G4cytosolic phospholipase A2

Background

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).
  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.

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FAQs for PLA2G4A (6659-PL). (Showing 1 - 1 of 1 FAQ).

  1. Hi. Actually I want to purchase anti cPLA2 antibody raised in rabbit which can bind to both phorphorylated and unphosphorylated mouse cPLA2 group IV. I want to a gel shift assay with western blotting to show that my protein of interest is causing the phosphorylation of cPLA2 inside the mouse macrophages RAW cells.Can you please tell me which antibody would be suitable for the purpose. Thanks a lot.
    • Here are our antibodies raised in rabbit that have been validated to detect cPLA2 in mouse samples in a western blot. We have not specifically confirmed the ability of these antibodies to detect both the phosphorylated and unphosphorylated forms of the protein, and I cannot guarantee that they will clearly differentiate between the two.

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Bioinformatics

Gene Symbol PLA2G4A
Uniprot