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IL-12 Protein, Mouse (HEK293, His)

Cat. No.: HY-P73159
COA Handling Instructions

Interleukin-12 subunit alpha (IL-12A; IL-12p35), an immune-suppressive cytokine, encodes a subunit of the cytokine IL-12 that acts on T and natural killer cells, and has a broad array of biological activities. IL-12A heterodimerizes with IL-12B to form the IL-12 cytokine or with EBI3/IL27B to form the IL-35 cytokine. IL-12 Protein, Mouse (HEK293, His) is produced in HEK293 cells with a C-Terminal His-tag. It consists of IL-12A (M1-A215) and IL-12B (M1-S335).

For research use only. We do not sell to patients.

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Description

Interleukin-12 subunit alpha (IL-12A; IL-12p35), an immune-suppressive cytokine, encodes a subunit of the cytokine IL-12 that acts on T and natural killer cells, and has a broad array of biological activities. IL-12A heterodimerizes with IL-12B to form the IL-12 cytokine or with EBI3/IL27B to form the IL-35 cytokine[1][2]. IL-12 Protein, Mouse (HEK293, His) is produced in HEK293 cells with a C-Terminal His-tag. It consists of IL-12A (M1-A215) and IL-12B (M1-S335).

Background

Interleukin-12 subunit alpha (IL-12A; IL-12p35), an immune-suppressive cytokine, encodes a subunit of the cytokine IL-12 that acts on T and natural killer cells, and has a broad array of biological activities. IL-12A is a disulfide-linked heterodimer composed of the 35-kD subunit encoded by this gene, and a 40-kD subunit that is a member of the cytokine receptor family. IL-12 is primarily produced by professional antigen-presenting cells (APCs) such as B-cells and dendritic cells (DCs) as well as macrophages and granulocytes, induces the production of IFN-gamma, favors the differentiation of Th1 cells and is an important link between innate resistance and adaptive immunity[1][2].
The amino acid sequence of human IL-12A protein has low homology between mouse IL-12A protein. While, human IL-12A shares 94% aa sequence identity with Rhesus Macaque IL-12A protein.
IL-12 family cytokines are pleiotropic immunological playmakers that coordinate innate and adaptive immune responses mainly via regulation of T-cell populations. The four core members of the Interleukin-12 (IL-12) family of cytokines, IL-12, IL-23, IL-27 and IL-35 are heterodimers which share α-cytokine subunits (IL-12p35 (IL-12A), IL-23p19, and IL-27p28) and β-cytokine subunits (IL-12p40, Ebi3). The subunits are each encoded by separate chromosomes and their expression is regulated independently. Among them, the IL-12A subunit has immunoregulatory functions hitherto attributed to IL-35. Pairing of the α-subunits, IL-12A or IL-23p19 with IL-12p40, gives rise to the two pro-inflammatory members IL-12 and IL-23, respectively, whereas the two immunosuppressive members of the family, IL-27 and IL-35, derive from pairing of IL-27p28 or IL-12A with Ebi3[1][2].
IL-12A suppresses lymphocyte proliferation, induces expansion of IL-10-expressing and IL-35-expressing B cells and ameliorates autoimmune uveitis in mice by antagonizing pathogenic Th17 responses. IL-12A-mediated expansion of Treg and Breg cells and its amelioration of experimental autoimmune encephalomyelitis (EAE) correlated with inhibition of cytokine-induced activation of STAT1/STAT3 pathways. IL-12A may be utilized for in vivo expansion of Tregs and Bregs cells and autologous Tregs and Bregs cell immunotherapy[1][2].

In Vitro

IL-12 alpha Protein, Mouse (6.25-1 ng/mL; 72 hours) suppresses T cell and B cell proliferation in dose-dependent manner[1].
IL-12 alpha Protein, Mouse (1 ng/mL; 1 hour) inhibits of cytokine-induced activation of STAT1/STAT3 pathways in CD4+ T cells. It also inhibits lymphocyte proliferation by suppressing the expressions of cell-cycle regulatory proteins in CD19+ B cells[2].

In Vivo

IL-12 alpha Protein, Mouse (1 ng/mouse; intraperitoneal injection; every other day; for 12 days) inhibits the expansion of pathogenic Th17 and Th1 cells and inhibits trafficking of inflammatory cells into the brain and spinal cord in mouse model of experimental autoimmune encephalomyelitis (EAE)[2].

Biological Activity

1.Measured by its ability to bind recombinant mouse IL12RB2 in a functional ELISA.
2.Measured by its ability to induce Interferon gamma secretion by human natural killer lymphoma NK-92 cells and the ED50 is 1-10 ng/mL.

Species

Mouse

Source

HEK293

Tag

C-6*His

Accession

P43431 (R23-A215) & P43432 (M23-S335)

Gene ID
Synonyms
IL-12; Interleukin 12; Interleukin-12 subunit alpha; IL-12A; Cytotoxic lymphocyte maturation factor 35 kDa subunit; CLMF p35; IL-12 subunit p35; Interleukin-12 subunit beta; IL-12B; Cytotoxic lymphocyte maturation factor 40 kDa subunit; CLMF p40; IL-12 subunit p40
Molecular Weight

Approximately 28&47 kDa

Purity

Greater than 90% as determined by reducing SDS-PAGE

Appearance

Lyophilized powder.

Formulation

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween 80 are added as protectants before lyophilization.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.

Storage & Stability

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Documentation
References
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

  • Reconstitution Calculator

  • Dilution Calculator

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The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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IL-12 Protein, Mouse (HEK293, His)
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HY-P73159
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