- Other Names
- Lipoprotein binding phospholipase A2 (Lp-PLA2), LDL-PLA2, Phospholipase A2 Group VII, platelet-activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) acetyl hydrolase (PAFAH), PAF-AH.
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- Product Citations
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Platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a phospholipid messenger synthesized by a variety of cells involved in host defense, such as endothelial cells, platelets, neutrophils, monocytes, and macrophages. High levels of PAF are responsible for a variety of human diseases such as inflammation, asthma, necrotizing enterocolitis, and sepsis. Phospholipase A2 Group VII (PLA2G7), also known as platelet- activating factor acetylhydrolase (PAFAH) or lipoprotein binding phospholipase A2 (Lp-PLA2), is an enzyme that hydrolyzed the acetyl group at the sn-2 position of the glycerol backbone from PAF to inactivate PAF. It has poor specificity toward Sn-2 long chain fatty acid. It also can hydrolyze oxidized phospholipids such as those within oxidized LDL, generating proinflammatory moieties lysophosphatidylcholine and oxidized fatty acids. Considering the proinflamatory properties of PAF and oxidized phospholipids, PLA2G7 plays a role in protecting cells from uncontrolled oxidative/inflammatory damage. A person with PLA2G7 deficiency is at an increased risk to develop a range of inflammatory disorders. PLA2G7 is produced primarily by macrophages and lymphocytes. The expression of plasma PLA2G7 is transcriptionally modulated by the mediators of inflammation, for example the promoter of the PLA2G7 gene is positively regulated by PAF and negatively regulated by interferon γ and lipopolysaccharide. The majority of the plasma PLA2G7 (approximately two-thirds) is bound to LDLs as it circulates in plasma, while the remaining plasma enzyme associates with HDLs and other lipoproteins.Product Details
- Human PLA2G7, amino acids Phe22-Asn441(Accession# BC038452) with a C-terminal TG-8H-GGQ tag was expressed in HEK 293E cells.
- Molecular Mass
- The 433 amino acid recombinant protein has a predicted molecular mass of approximately 49.3 kD. The protein migrates at approximately 60 kD in DTT-reducing conditions and at approximately 50 kD in non-reducing conditions by SDS-PAGE.
- >90%, as determined by Coomasie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in pH 5.5 buffer containing 25 mM MES and 150 mM NaCl.
- Endotoxin Level
- Less than 1.0 EU per µg of protein as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.) Please note, new lots of the 100 µg size will be lot-specific and may differ from previous lots that had a fixed concentration.
- Storage & Handling
- Unopened vial can be stored at -20°C or -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.
- Human PLA2G7 hydrolyzes a chromogenic substrate, 2-thio-PAF (1-O-hexadecyl-2-deoxy-2-thio-S-acetyl-sn-glyceryl-3-phosphorylcholine), with a specific activity value higher than 14000 pmol/µg/min.
- Recommended Usage
- Application Notes
Human PLA2G7/Lp-PLA2 Activity Assay
Human PLA2G7 activity is measured by its ability to cleave a chromogenic peptide substrate 2-Thio-PAF. The progress of the product formation is monitored by increase in intensity of absorbance at 405 nm in the presence of DTNB.
Materials and Buffers
1. Assay Buffer: pH 7.0, 25 mM Sodium Phosphate, 150 mM NaCl, 0.1% BSA (w/v)
2. Human PLA2G7
3. PLA2G7 substrate: 2-Thio-PAF (1-O-hexadecyl-2-deoxy-2-thio-S-acetyl-sn-glyceryl-3-phosphorylcholine), 20 mM in DMSO
4. DTNB (5,5'-dithio-bis(2-nitrobenzoic acid)), 20 mM in DMSO
1. Dilute activated hPLA2G7 to 0.2 µg/mL with Assay Buffer.
2. Dilute the substrate and DTNB together at 200 µM each in Assay Buffer.
3. Load 50 µL of the diluted hPLA2G7 solution into a plate and 50 µL of assay buffer for the substrate blank, and start a reaction by adding 50 µL of 200 µM of the substrate stock solution into each well.
4. Read the reaction progress by monitoring absorbance at 405 nm in kinetic mode for five minutes.
5. The final human hPLA2G7 concentration is at 0.1 µg/mL (0.01 µg), the substrate concentration at 100 µM, and DTNB at 100 µM.
Conversion factor at 405 nm is 23.5 pmol/mAu by using extinction coefficient 13260 M-1cm-1 and 0.32 cm of light path correction at total volume of 100 µL.
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at email@example.com.
(PubMed link indicates BioLegend citation)
- Al-Darmaki S, et al. 2003. J. Immunol. 170:167.
- Silva IT, et al. 2011. Lipids Health Dis. 10:170.
- Marathe GK, et al. 2014. J. Lipid Res. 55:1847.
- Tselepis AD. 2016. J. Biomed. Res. 31:1.
- Human serum and plasma, high expression in U-251 cell line and medium expression in U-2 and A-451 cell line.
- Platelet-activating factor (PAF) inactivation.
- LDL, HDL, VLDL.
- Inactivate platelet-activating factor and hydrolyze oxidized phospholipids.
- Molecular Family
- Enzymes and Regulators, Phospho-Proteins
- Gene ID
- 7941 View all products for this Gene ID
- View information about PLA2G7 on UniProt.org
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.