Recombinant Human IL-12 (p70) (293E expressed, carrier-free)

Product Details

Source

Human IL-12 consists of two subunits IL-12B (p40) amino acid (Ile23- Ser328, Accession: # P29460) fusion with human IL-12A (p35) amino acids (Arg23-Ser219, Accession: #P29459) with a C-terminus 6His was expressed in 293E cells.

Molecular Mass

The 332 amino acid recombinant protein has a predicted molecular mass of approximately 59.8 kD. The DTT-reduced and non-reduced protein migrates at approximately 65 -75 kD and 62 - 70 kD by SDS-PAGE respectively. The predicted N-terminal amino acid is Ile.

Purity

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

Formulation

0.2 µm filtered protein solution is in PBS, pH 7.2

Endotoxin Level

Less than 0.1 EU per µg of protein 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 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 the 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% endotoxin-free 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 or stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.

Activity

Human IL-12 induces IFN-γ production in PHA activated human PBMC in a dose-dependent manner. The ED₅₀ for this effect is 0.009 – 0.09 ng/mL.

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 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

IL-12 is produced by monocytes, macrophages, neutrophils, dendritic cells and B cells. In the CNS, astrocytes and microglia are the main sources of IL-12.

Function

IL-12 is produced by myeloid cells and DCs in response to microbial stimuli, such as those mediated by bacteria, fungi, viruses, and intracellular parasites. IL-12 drives Th1 differentiation and IFN-γ production. IL-12 acts as a bridge between innate resistance and adaptive immunity. IL-12 production by activated APC is suppressed by IL-10. In addition, IL-12 production by macrophages is regulated by TNFα and nitric oxide. TLR-4 and TLR-9 can cooperate to increase selectively IL-12 production by murine dendritic cells.

Interaction

Cells of hematopoietic origin express the IL-12R, including NK cells, activated T-cells and dendritic cells.

Ligand/Receptor

IL-12 receptor is a heterodimer containing IL-12Rβ1 and IL-12Rβ2 subunits.

Bioactivity

Measured by its ability to induce IFN-γ production in PHA activated human PBMC

Biology Area

Immunology, Innate Immunity

Molecular Family

Cytokines/Chemokines

Antigen References

1. Schoenhaut DS, et al. 1992. J Immunol. 148:3433-40.
2. Manetti R, et al. 1994. J Exp Med. 179:1273-83.
3. Ireland D, et al. 2005. Viral Immunol. 18:397-402.
4. Moreno SE, et al. 2006. J Immunol. 177:3218-24.
5. Lyakh L, et al. 2008. Immunol Rev. 226:112-31.
6. Theiner G, et al. 2008. Mol Immunol. 45:244-52.
7. Zhu S, et al. 2010. J Immunol. 184:2348-54.
8. Nguyen KG, et al 2020. Front Immunol. 11:575597.

Gene ID

3593 View all products for this Gene ID 3592 View all products for this Gene ID

UniProt

View information about IL-12 on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Material 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