Recombinant Mouse CD39 (carrier-free)

Product Details

Source

Mouse CD39 amino acids Thr38 - Ile478 (Accession # P55772) was expressed in CHO cells.The carboxy terminus contains TG-8His-GGQ tag.

Molecular Mass

The 454 amino acid recombinant protein has a predicted molecular mass of approximately 51.1 kD. The DTT-reduced and non-reduced protein migrate at approximately 80 – 150 kD and 100-200 kD respectively by SDS-PAGE. The predicted amino terminal is Thr.

Purity

> 95% by SDS-PAGE gel as determined by Coomassie stained SDS-PAGE.

Formulation

0.22 µm filtered protein solution is in 25mM Tris pH 7.20, 10mM CaCl₂, 150mM NaCl, 10% Glycerol.

Endotoxin Level

Less than 0.10 EU per µg cytokine 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

The activity of mouse CD39 is measured by its property to hydrolyze the 5’- phosphate groups from the substrate adenosine-5’-triphosphate (ATP). The specific activity is >25,000 pmol/min/µg.

Application

Bioassay

Application Notes

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.
 

Mouse CD39 Activity assay

 

Solutions:

Enzyme: Recombinant mouse CD39

Substrate: Adenosine triphosphate (ATP), 10 uM stock solution in deionized water.

Assay buffer: 25 mM Tris, 5 mM CaCl₂, pH 7.5

Malachite Green Phosphate Assay Kit

Procedure

1. Phosphate Standard curve. From a 1 M Phosphate Standard solution (included in the Malachite Green Phosphate Assay Kit) prepare 100 µM solution of the standard in the assay buffer (phosphate stock). Set six points of 1:2 serial dilutions for the standard curve from 2.5 to 0.39 nmol/well.
2. Prepare (1:2) serial dilutions of recombinant mouse CD39 in Assay Buffer. The top concentration will be 0.5 ng/well in 25 µl.
3. Prepare the substrate (ATP) solution at 100 µM and transfer 25 µl to each well containing the recombinant mouse CD39 and the blanks (buffer only).
4. Incubate the plate at 37°C for 30 minutes.
5. Add 5 ul of acidic solution (included in the Malachite Green Phosphate Assay Kit) to all the wells, mix gently and incubate for 10 minutes at room temperature.
6. Add 15 µl of Malachite Green Blue solution to each well. Mix gently and incubate 20 minutes at Room temperature.
7. Measure the absorbance using a microplate reader set at 620 nm.

Antigen Details

Structure

Monomer

Distribution

Activated lymphoid cells, activated B cells, endothelial cells, NK cells, dendritic cells, Langerhans cells, monocytes, macrophages, mesangial cells, neutrophils, CD4 T cells, regulatory T cells, and CD8+ cytotoxic T cells.

Function

Hydrolyzes extracellular ATP and less efficiently ADP to AMP (Ca²⁺, Mg²⁺ dependent). Regulated by pro-inflammatory cytokines (e.g. IL-6 and IL-27), oxidative stress, and hypoxia.

Ligand/Receptor

ATP and ADP

Bioactivity

Hydrolyzes extracellular ATP and less efficiently ADP to AMP

Biology Area

Immunology, Innate Immunity

Molecular Family

Enzymes and Regulators, Phospho-Proteins, CD Molecules

Antigen References

1. Maliszewski CR, et al. 1994. J Immunol. 153:3574.
2. Heine P, et al. 2001. Eur J Biochem. 268:364.
3. Grinthal A, Guidotti G. 2006. Purinergic Signal. 2:391.
4. Köhler D, et al. 2007. Circulation. 116:1784.
5. Eltzschig HK, et al. 2009. Blood. 113:224.
6. Zebisch M, et al. 2012 J Mol Biol. 415:288.
7. Yegutkin GG, et al. 2012. FASEB J. 26:3875.
8. Wheeler DG, et al. 2012. J Mol Cell Cardiol. 52:958.
9. Antonioli L, et al. 2013. Trends Mol Med. 19:355.

Gene ID

12495 View all products for this Gene ID

UniProt

View information about CD39 on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• 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