Cell-Vive™ GMP Recombinant Human IL-12 (p70) (carrier-free)

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Other Names
Natural killer cell stimulatory factor (NKSF), cytotoxic lymphocyte maturation factor (CLMF)
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Human_IL-12_CF_GMP-RECOM_042222
GMP recombinant human IL-12 induces IFN-γ production in PHA activated human PBMC in a dose-dependent manner with ED50 range of 0.01 - 0.1 ng/mL.
  • Human_IL-12_CF_GMP-RECOM_042222
    GMP recombinant human IL-12 induces IFN-γ production in PHA activated human PBMC in a dose-dependent manner with ED50 range of 0.01 - 0.1 ng/mL.
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573014 25 µg £702
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573016 100 µg £2245
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Description

IL-12 (p70) is a disulfide-linked heterodimer composed of unrelated p40 (glycosylated) and p35 subunits. IL-12 acts as a growth factor for activated human T and NK cells, enhances the lytic activity of human NK cells, and stimulates the production of IFNg by resting human PBMC. IL-12R is formed by two chains, IL-12Rβ1 and IL-12Rβ2. IL-12Rβ1 is associated with the Janus kinase (Jak) Tyk2 and binds IL-12 p40; IL-12Rβ2 is associated with Jak2 and binds either the heterodimer or the p35 chain. Signaling through the IL-12 receptor complex induces phosphorylation, dimerization, and nuclear translocation of several signal transducers and activators of transcription (STAT) family members (STAT1, 3, 4, 5), but most of the biological responses to IL-12 have been attributed to STAT4. IL-12 has been shown to elicit anti-tumor activity in mice and humans. It is believed that the antitumor effects of IL-12 are mediated, at least in part, by indirect mechanisms. Induction of IFN-γ results in the upregulation of class I and class II MHC molecules, adhesion molecules (ICAM-1), nitric oxide production by antigen presenting cells (APC), and the production of additional cytokines, CXCL9 and 10, which in turn mediate angiostatic effects. Cytokine detection IL-12, IL-23 and IL-35 share common subunits, utilizing combinations of p40, p19 and p35 proteins. Caution must be used when selecting antibodies and assays when specific identification, measurement, as well as activation state discrimination, is required.

Product Details
Quality Statement

BioLegend Cell-Vive™ GMP Recombinant proteins are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Ph. Eur. Chapter 5.2.12 in a dedicated GMP facility compliant with ISO 13485:2016. Specifications and processes include:

  • Low endotoxin level (≤0.1 EU/μg)
  • Purity (≥ 95% or higher)
  • Bioburden testing
  • Mycoplasma testing
  • Batch-to-batch consistency
  • Vendor qualification
  • Raw material traceability and documentation
  • Documented procedures and employee training
  • Equipment maintenance and monitoring records
  • Lot-specific certificates of analysis
  • Quality audits per ISO 13485:2016
  • QA review of released products
Technical Data Sheet (pdf)

Product Details

Source
Expressed in insect cells as secreted protein (p35: Accession# NM_000882, p40: Accession# NM_002187)
Molecular Mass
The hIL-12 consists of two subunits linked via a disulfide bond: P35 (Accession# NP_000873.2: Arg 57- Ser 253) and P40 (Accession# NP_002178.2: Ile 23-Ser 328). The total predicted molecular weight is 57 kDa. The non-reduced protein migrates at approximately 55 kDa and the DTT-reduced protein produces two bands migrating at approximately 26 kDa and 40 KDa by SDS-PAGE.
N-terminal
Sequence Analysis

p40 Subunit: Ile-Trp-Glu-Leu-Lys-Lys-Asp-Val-Tyr-Val

p35 Subunit: Arg-Asn-Leu-Pro-Val-Ala-Thr-Pro-Asp-Pro

Purity
≥ 95%, as determined by Coomassie stained SDS-PAGE
Formulation
0.1 µm filtered protein solution is in PBS, pH 7.2.
Endotoxin Level
Less than or equal to 0.1 EU per μg protein as determined by the LAL method
Concentration
500 µg/mL
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% 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
ED50 = 0.01 - 0.1 ng/mL as determined by the dose-dependent production of IFN-γ in PHA activated human PBMC. The specific activity of Cell-Vive™ GMP Recombinant Human IL-12 (carrier-free) is ≥ 2.8 x 106 IU/mg when compared against the WHO International Standard for human IL-12 (NIBSC code: 95/544).
Application

Bioassay

Cell culture

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 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.

Disclaimer

BioLegend Cell-Vive™ GMP Recombinant proteins are for research use only. Suitable for ex vivo cell processing. Not for injection or diagnostic or therapeutic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

Antigen Details

Structure
Cytokine
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 IFNγ 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.
Gene ID
3592 View all products for this Gene ID 3593 View all products for this Gene ID
UniProt
View information about IL-12 on UniProt.org
Go To Top Version: 2    Revision Date: 08/26/2024

For Research Use Only. Suitable for ex vivo cell processing. Not for injection or diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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