IL-12 Protein, Rhesus Macaque (HEK293, His)

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, Rhesus Macaque (HEK293, His) is produced in HEK293 cells with a C-Terminal His-tag . It consists of IL-12A (M1-S219) and IL-12B (M1-S328).

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].

IL-12 Protein, Human (1 ng/mL; for 5 days) results in the induction of IFN-γ secretion by Dermatophagoides pteronyssinus group 1 antigen (Der p 1)-specific CD4+ T cells in dendritic cell (DC)-T cell cocultures, whereas their production of IL-5 is not inhibited^[3].

IL-12 Protein, Rhesus Macaque (1 and 2 μg/kg; s.c.; twice a week; for 8 weeks) significantly protects macaques from SIVmac251-induced disease. The higher IL-12 dose leads to lower plasma viral loads and markedly lower peripheral blood mononuclear cell and lymph node proviral DNA loads^[4].

Measured by its ability to induce Interferon gamma secretion by human natural killer lymphoma NK-92 cells. The ED₅₀ for this effect is typically 0.2-2ng/mL.

Rhesus Macaque

HEK293

C-His

P48091 (R23-S219)&P48095 (I23-S328)

703205  [NCBI]&694747  [NCBI]

IL12A:RNLSVATPGP
EMFPCLHHSQNLLKAASNTLQKARQILEFYPCTSEEIDHEDITKDKTSTVEACLPLELIKNESCLNSRETSFITNGSCLASRKTSFMMALCLRSIYEDLKMYQVEFKTMNAKLLRDPKRQIFLDQNILGVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRV
MSYLNAS <br/>IL12B:IWELKKDVYV
VELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSGEVLGSGKTLTIQVKEFGDAGQYTCHKGGEALSHSLLLLHKKEDGIWSTDVLKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSNPQGVTCGAVTLSAERVRGDNKEYEYSVECQEDSACPAAEERLPIEVMVDAIHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCIQVQGKSKREKKDRIFTDKTSATVICRKNASFSVQAQDRYYSSSWSEWASVPCS

Approximately 60.2 kDa

Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder

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.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

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.

Room temperature in continental US; may vary elsewhere.

• [1]. Ivy M Dambuza, et al. IL-12p35 induces expansion of IL-10 and IL-35-expressing regulatory B cells and ameliorates autoimmune disease. Nat Commun. 2017 Sep 28;8(1):719.  [Content Brief]

  [2]. Jin Kyeong Choi, et al. IL-12p35 Inhibits Neuroinflammation and Ameliorates Autoimmune Encephalomyelitis. Front Immunol. 2017 Oct 5;8:1258.  [Content Brief]

  [3]. D De Wit, et al. Helper T-cell responses elicited by Der p 1-pulsed dendritic cells and recombinant IL-12 in atopic and healthy subjects. J Allergy Clin Immunol. 2000 Feb;105(2 Pt 1):346-52.  [Content Brief]

  [4]. A A Ansari, et al. Administration of recombinant rhesus interleukin-12 during acute simian immunodeficiency virus (SIV) infection leads to decreased viral loads associated with prolonged survival in SIVmac251-infected rhesus macaques. J Virol. 2002 Feb;76(4):1731-43.  [Content Brief]