Recombinant Mouse DPP4 (carrier-free)

Product Details

Source

Mouse DPP4, amino acid (Ser29 – His760) (Accession: BC022183) was expressed in CHO cells. The C-terminus contains TG-10His-GGQ tag.

Molecular Mass

The 747 amino acid recombinant protein has a predicted molecular mass of approximately 86.2 kD. The DTT-reduced and non reduced proteins migrate at approximately 90 - 100 kD by SDS-PAGE. The carboxi-terminus contains TG-10His-GGQ – tag. The predicted N-terminal amino acid is Ser.

Purity

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

Formulation

0.22 µm filtered protein solution is in 20 mM TRIS, 300 mM NaCl, pH 7.5.

Endotoxin Level

Less than 0.1 EU per µg cytokine as determined by the LAL method

Concentration

25 µg size is 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 -70ᵒC for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.

Activity

Mouse DPP4 activity is measured by its ability to cleave a fluorogenic peptide substrate Gly-Pro-AMC. The increase of the product is monitored by increase in intensity of fluorescence at 460 nm with excitation at 380 nm. The specific activity is > 6000 pmol/min/µg in the presence of 0.005 µg of recombinant mouse DPP4.

Application

Bioassay

Application Notes

Mouse DPP4 Activity Assay

 

Mouse DPP4 activity is measured by its ability to cleave a fluorogenic peptide substrate Gly-Pro-AMC. The increase of the product is monitored by increase in intensity of fluorescence at 460 nm with excitation at 380 nm. This protein is in the activated form and does not require further activation for bioassay. The purified recombinant DPP4 behaves as a non-disulfide-bonded dimer based on the analysis of SDS PAGE and gel filtration.

 

Materials

1. Assay buffer:  pH 8.0, 25 mM Tris

2. Recombinant mouse DPP-4

3. DPP-4 substrate: Gly-Pro-AMC

 

Activity assay procedures

1. Dilute the mouse DPP4 in the assay buffer at 0.1 µg/mL (0.1 ng/µL).

2. Dilute the substrate at 100 µM in assay buffer.

3. Load into a well plate 50 μL of the 0.1 ng/μL mouse DPP4 and start the reaction by adding 50 μL of 100 μM substrate. Include a substrate blank containing 50 μL of assay buffer and 50 μL of 100 μM substrate solution.

4. Read the product formation by measuring 380/460 nm (Excitation/Emission) in kinetic mode for 5 minutes.

 

The final mouse DPP4 concentration is 0.05 µg/mL (0.05 ng/uL; 5 ng)

The final concentration of the substrate is 50 µM.

 

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 tech@biolegend.com.

Antigen Details

Structure

Monomer

Distribution

Ubiquitously in many tissues (mainly expressed in large intestine), T cells, adipocytes

Function

Regulates of T-cell activation, upregulating CD86 expression and NF-κB pathway, lymphocyte-epithelial cell adhesion. Regulates postprandial glucose through degradation of GLP-1. Proteolysis of the extracellular matrix, removes amino-terminal dipeptides.

Interaction

T cells, lymphocytes

Ligand/Receptor

Adenosin deaminase (ADA), RANTES, stromal cell-derived factor (SDF)-1α, glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide-1 (GLP-1)

Bioactivity

Mouse DPP4 activity is measured by its ability to cleave a fluorogenic peptide substrate Gly-Pro-AMC.

Cell Type

Endothelial cells, Lymphocytes, T cells

Biology Area

Cell Biology, Costimulatory Molecules, Immunology

Molecular Family

Adhesion Molecules, Enzymes and Regulators, Proteases

Antigen References

1. Morimoto C, et al. 1989. J Immunol. 143:3430-9.
2. Engel M, et al. 2003. Proc Natl Acad Sci U S A. 100:5063-8.
3. Ohnuma K, et al. 2007. J Biol Chem. 282:10117-10131.
4. Wang N, et al. 2013. Cell Res. 23:986-93.
5. Song W, et al. 2014. Virology. 471-473:49-53.
6. Metzemaekers M, et al. 2016. Front Immunol. 7:483.
7. Varin EM, et al. 2019. Cell Metab. 29:320-334.
8. Hartenian E. et al. 2020. J Biol Chem. 295:12910-12934.

Gene ID

13482 View all products for this Gene ID

UniProt

View information about DPP4 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