Recombinant Human PLA2G7 (carrier-free)

Product Details

Source

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.

Purity

>90%, as determined by Coomasie stained SDS-PAGE.

Formulation

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.

Concentration

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. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.

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.

Activity

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

Bioassay

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

Assay Procedures

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 verified 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 tech@biolegend.com.

Application References

(PubMed link indicates BioLegend citation)

1. Al-Darmaki S, et al. 2003. J. Immunol. 170:167.
2. Silva IT, et al. 2011. Lipids Health Dis. 10:170.
3. Marathe GK, et al. 2014. J. Lipid Res. 55:1847.
4. Tselepis AD. 2016. J. Biomed. Res. 31:1.

Antigen Details

Structure

Monomer

Distribution

Human serum and plasma, high expression in U-251 cell line and medium expression in U-2 and A-451 cell line.

Function

Platelet-activating factor (PAF) inactivation.

Ligand/Receptor

LDL, HDL, VLDL.

Bioactivity

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

UniProt

View information about PLA2G7 on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Pages & Pathways

Related FAQs

Why choose BioLegend recombinant proteins?

     • Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
     • Greater than 95% Purity or higher, tested on every lot of product.
     • 100% Satisfaction Guarantee for quality performance, stability, and consistency.
     • Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
     • Bulk and customization available. Contact us.
     • Learn more about our Recombinant Proteins.

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.

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.

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.

How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?

Use formula Specific activity (Units/mg) = 10^6/ ED50 (ng/mL)

Certificate of Analysis