Secretory Phopholipase A2 is an enzyme that hydrolyses the sn-2 ester bond of phospholipids and cell membranes, generating non-esterified free fatty acids and lysophospholipids (1‑3). Most secretory PLA2s are stored in cytoplasmic granules and are released in the extracellular environment on appropriate cell activation. Thus, they are present at higher concentration in the plasma and biological fluids of patients with systemic inflammatory, autoimmune, or allergic disease, such as acute pancreatitis, rheumatoid arthritis, bronchial asthma, and allergic rhinitis. PLA2G1B has been thought to play major role in digestion of glycerophospholipids in nutrients, given its abundance in digestive organs (4). Since its expression has been observed in non-digestive organs including the lung, spleen, kidney, ovary, retina, brain, and neurons, its function may not limited to digestive role (5, 6). rhPLA2G1B has been maturated by cleaving the pro-peptide with trypsin, and the mature active form was further purified with ion-exchange column chromatograpy.
Recombinant Human PLA2G1B Protein, CF
R&D Systems | Catalog # 5018-PL
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Key Product Details
- R&D Systems NS0-derived Recombinant Human PLA2G1B Protein (5018-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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human PLA2G1B protein
Ala23-Ser148, with a C-terminal 6-His tag
Ala23-Ser148, with a C-terminal 6-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
Ala23
Predicted Molecular Mass
15 kDa
SDS-PAGE
18 kDa, reducing conditions
Activity
Measured by its ability to hydrolyze 1-Hexadecanoyl-2-(1-pyrene-decanoyl)-sn-glycero-3-phosphoglycerol.
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
5018-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.
|
Background: PLA2G1B
References
- Webb, N. R. (2005) Cur. Opin. Lipid. 16:341.
- Triggiani, M. et al. (2005) J. Allergy Clin. Immunol. 116:1000.
- Murakami, M. and Kudo, I. (2004) Biol. Pharm. Bull. 27:1158.
- de Haas, G. H. et al. (1968) Biochime. Biophysic. Acta. 159:118.
- Kolko, M. et al. (2005) Cell. Mol. Neurobiol. 25:1107.
- Kolko, M. et al. (2007) Acta Ophthalmol. Scand. 85:317.
Long Name
Phospholipase A2 Group IB
Alternate Names
PLA2, PLA2A, PPLA2
Gene Symbol
PLA2G1B
UniProt
Additional PLA2G1B Products
Product Documents for Recombinant Human PLA2G1B 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.
Product Specific Notices for Recombinant Human PLA2G1B Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human PLA2G1B Protein, CF (5018-PL):
Materials
- Assay Buffer: 50 mM HEPES, 1 mg/mL BSA, pH 7.0.
- Substrate Buffer: 50 mM HEPES, 20 mM CaCl2, 1 mg/mL BSA, pH 7.0.
- Recombinant Human Phospholipase A2 Group IB/PLA2G1B (rhPLA2G1B) (Catalog # 5018-PL)
- Substrate: 1-hexadecanoyl-2-(1-pyrene-decanoyl)-sn-glycero-3-phosphoglycerol Ammonium Salt, 2 mM stock in DMSO
- Black 96-well Plate
- Fluorescent Plate Reader
- Thaw Substrate at 37 °C for five minutes. Mix well.
- Dilute rhPLA2G1B to 0.02 ng/µL in Assay Buffer.
- Dilute Substrate to 100 µM in Substrate Buffer.
- Load into plate, 50 µL of 0.02 ng/µL rhPLA2G1B, and start reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
- Read at excitation and emission wavelengths of 345 nm and 395 nm (top read), respectively in kinetic mode for 5 minutes. Shake the plate for 3 seconds between each read.
- 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.
- rhPLA2G1B: 0.001 µg
- Substrate: 50 µM