Recombinant Human Osteopontin (carrier-free)

Product Details

Source

Human Osteopontin, amino acids (Ile17-Asn300) (Accession# NP_000573.1), was expressed in 293E cells with a C-terminal His tag.

Molecular Mass

The 290 amino acid recombinant protein has a predicted molecular mass of approximately 33 kD. The DTT-reduced and non-reduced protein migrates approximately 55 kD by SDS-PAGE. The predicted N-terminal amino acid is Ile.

Purity

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

Formulation

0.22 µm filtered protein solution is in PBS, pH 7.2.

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 between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.

Activity

Human osteopontin inhibits the production of IFN-γ in human activated T cells in a dose-dependent manner. The ED₅₀ for this effect is 0.2 – 2.0 ng/mL.

Application

Bioassay

Application Notes

Bioassay Protocol¹

1. Increasing concentrations of hOsteopontin, along wtih 500 ng/ml of anti-CD3 antibody (OKT3), were used to coat a 96-well plate overnight.
2. Excess hOsteopontin and OKT3 were washed away with PBS before incubating purified human T cells at 2 X 10⁵ cells per well.
3. The supernatant cell culture was collected after 24 - 48 hours of incubation and the concentration of IFN-γ was detected using BioLegend's Human IFN-γ ELISA MAX™ Deluxe (Cat No: 430104).

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. O'Regan AW, et al. 2000. J. Leuko. Biol. 68:495.

Product Citations

1. Anchan A, et al. 2020. Int J Mol Sci. :21. PubMed

Antigen Details

Structure

Monomer.

Distribution

Osteoblasts, osteoclasts, endothelial cells, macrophages, and smooth muscle cells.

Function

Plays a role in cell migration, survival, inhibition of calcification, and regulation of immune cell function. Hypocalcemia and hypophosphatemia leads to an increase in Osteopontin transcription, translation, and secretion.

Interaction

T cells, neutrophils, macrophages, osteoclasts, and mast cells.

Ligand/Receptor

CD44, integrins (αVβ3).

Bioactivity

Human OPN1 inhibits IFN-γ secretion by anti-CD3 activated human T cells.

Biology Area

Apoptosis/Tumor Suppressors/Cell Death, Cancer Biomarkers, Cell Biology, Cell Motility/Cytoskeleton/Structure, Immunology, Signal Transduction, Stem Cells

Molecular Family

Phospho-Proteins

Antigen References

1. Wang KX and Denhardt DT. 2008. Cytokine Growth Factor Rev. 333.
2. Agnihotri R, et al. 2001. J. Biol. Chem. 276:28261.
3. O'Regan AW, et al. 2000. J. Leuko. Biol. 68:495.
4. Yokosaki Y, et al. 2005. Matrix Biol. 24:418.
5. Rangaswami H, et al. 2006. Trends Cell Biol. 16:79.

Gene ID

6696 View all products for this Gene ID

UniProt

View information about Osteopontin 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