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  6. GMP IL-12 Protein, Human (HEK293)

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. GMP IL-12 Protein, Human (HEK293), a recombinant GMP-grade protein,  is produced in HEK293 cells. It consists of IL-12A and IL-12B.

GMP-grade recombinant proteins are produced under independent QA supervision, as well as quality records and full traceability.

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]. GMP IL-12 Protein, Human (HEK293), a recombinant GMP-grade protein,  is produced in HEK293 cells. It consists of IL-12A and IL-12B.

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 Protein, Human (10 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].

Biological Activity

Immobilized Human IL-12RB1-His at 10 μg/mL (100 μl/well)can bind Human IL-12*: Biotinylated by NHS-biotin prior to testing. The ED50 for this effect is 7 μg/mL.

Species

Human

Source

HEK293

Tag

Tag Free

Accession

P29459 (R23-S219) & P29460 (I23-S328)

Gene ID

3592  [NCBI]&3593  [NCBI]

Molecular Construction
N-term
IL12A (R23-S219)
Accession # P29459
C-term
N-term
P29460 IL12B (I23-S328)
Accession #
C-term
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
AA Sequence

RNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS
&:
IWELKKDVYVVELDWYPDAPGEMVVLTCDTPE

Molecular Weight

25-38&40-50 kDa

Purity

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

Appearance

Lyophilized powder.

Formulation

Lyophilized from a 0.2 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.

Endotoxin Level

<0.1 EU/μg, determined by LAL method.

Reconstitution

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

Storage & Stability

Stored at -20°C for 2 years from date of receipt. 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
  • How should lyophilized recombinant proteins be reconstituted and stored?

    1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss. 2. Taking 10 μg as an example, first add 20 μL of reconstituted solution provided by MCE and use a pipette to gently resuspend the lyophilized protein until it is fully dissolved.. (For most proteins, the reconstitution solution we provide is sterile water. If a diluent other than water is required, it will be indicated in the product's Certificate of Analysis (COA).). 3. Add an additional 80 μL of buffer/culture medium containing carrier protein (either 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose), and then use a pipette to gently mix until uniform. The final concentration is should not be lower than 100 μg/mL. 4. Aliquot at least 20 μL per tube. 5. After aliquoting, store it frozen at a temperature ranging from -20ºC to -80ºC, and it can be preserved for 3 to 6 months.

  • How should solution-form recombinant proteins be stored?

    1. The product can be stored in its original form and diluted as needed upon use. 2. Alternatively, dilute with a buffer/culture medium containing a carrier protein (either 0.1% BSA, 5% HSA, 10% FBS, or 5% alginate), mix well by pipetting, and ensure that

  • Why is it necessary to add carrier proteins?

    Carrier proteins are commonly added to enhance the stability of recombinant proteins, preventing them from adhering to the walls of the container during freezing or thawing processes. Plastic tubes have a certain adsorptive capacity for proteins, which may lead to difficulty in separating the protein from the tube walls, resulting in a decrease in the actual concentration of the protein in the solution and thus affecting its activity. To minimize such losses, it is recommended to add a commonly used carrier protein solution prior to the long-term storage of recombinant protein products.

  • Carrier protein types and options?

    In cases where the carrier protein is not expected to influence the experimental outcomes, an appropriate carrier protein, such as 0.1% BSA (Bovine Serum Albumin), 5% HSA (Human Serum Albumin), 10% FBS (Fetal Bovine Serum), or 5% trehalose, can be incorpo

  • Reconstitution Calculator

  • Dilution Calculator

  • Specific Activity Calculator

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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Product Name:
GMP IL-12 Protein, Human (HEK293)
Cat. No.:
HY-P7032G
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