Recombinant Human PLA2G1B Protein, CF

Catalog #: 5018-PL Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
5018-PL-010

R&D Systems Recombinant Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human PLA2G1B Protein, CF Summary

Product Specifications

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.

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.

Source

Mouse myeloma cell line, NS0-derived human PLA2G1B protein
Ala23-Ser148, with a C-terminal 6-His tag

Accession #

NP_000919

N-terminal Sequence
Analysis

Ala23

Predicted Molecular Mass

15 kDa

SDS-PAGE

18 kDa, reducing conditions

Product Datasheets

5018-PL

Product Datasheet

COA

SDS

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.

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. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

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 V_max* (RFU/min) x Conversion Factor** (pmol/RFU)
	amount of enzyme (µg)

*Adjusted for Substrate Blank
**Derived using calibration standard 1-pyrenedecanoic acid.

Per Well:

rhPLA2G1B: 0.001 µg
Substrate: 50 µM

Reconstitution Calculator

Background: PLA2G1B

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.

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

Entrez Gene IDs

5319 (Human); 18778 (Mouse); 29526 (Rat)

Alternate Names

Group IB phospholipase A2; Phosphatidylcholine 2-acylhydrolase 1B; phospholipase A2; phospholipase A2, group IB (pancreas); PLA2; PLA2A; PLA2AMGC119834; PLA2EC 3.1.1.4; PLA2G1B; PPLA2; PPLA2MGC119835