What is the difference between Recombinant Human IL-12 Protein (Catalog # 219-IL) and Recombinant Human Il-12 (linked heterodimer) Protein (Catalog # 10018-IL?

Name: Recombinant Human IL-12 (linked heterodimer) Protein, CF
Brand: R&D Systems
SKU: 10018-IL

The Recombinant Human IL-12 Protein (Catalog # 219-IL), expressed in Sf 21 cells, is a disulfide-linked homodimer consisting of the p35 and p40 subunits. The subunits can be separated under reducing conditions. The Recombinant Human IL-12 (linked heterodimer) Protein (Catalog # 10018-IL), expressed in HEK293 cells, is a chimeric protein consisting of the p35 and p40 subunits connected by a linker. These two subunits cannot be separated.

Recombinant Human IL-12 (linked heterodimer) Protein, CF

Catalog # 10018-IL

Product Datasheet / COA / SDS

GS Linker, Defined heterodimer stoichiometry.

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Key Product Details

Source:	HEK293
Accession #:	P29460 (IL-12p40), P29459 (IL-12p35)
Structure / Form:	GS-linked heterodimer
Applications:	Bioactivity

View Full Technical Details for Recombinant Human IL-12 (linked heterodimer) Protein, CF

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human IL-12 protein

Human IL-12p40 (Ile23-Ser328) Accession # P29460	GSGSSRGGSGSGGSGGGGSK	Human IL-12p35 (Arg23-Ser219) Accession # P29459
N-terminus		C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal sequence Analysis

Ile23

Predicted Molecular Mass

59 kDa

SDS-PAGE

63-75 kDa, reducing conditions

Activity

Measured in a cell proliferation assay using PHA-stimulated human T lymphoblasts. Symons, J.A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 272.

The ED₅₀ for this effect is 0.01-0.05 ng/mL.

Recombinant Human IL-12 (linked heterodimer) Protein, CF Scientific Data Examples

	Recombinant Human IL-12 (linked heterodimer) Protein Bioactivity Recombinant Human IL-12 (Catalog # 10018-IL) stimulates proliferation in PHA-activated human T lymphoblasts. The ED₅₀ for this effect is 0.01-0.05 ng/mL.
	Recombinant Human IL-12 (linked heterodimer) Protein SDS-PAGE 2 μg/lane of Recombinant Human IL‑12 was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 63-75 kDa.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10018-IL

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100-200 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, ≤ -20 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IL-12

Interleukin 12, also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a pleiotropic cytokine originally identified in the medium of activated human B lymphoblastoid cell lines (1). The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the human IL-6 receptor while the p35 subunit shows distant but significant sequence similarity to IL-6, G-CSF, and chicken MGF (2, 3). These observations have led to the suggestion that IL-12 might have evolved from a cytokine/soluble receptor complex. Human and murine IL-12 share 70% and 60% amino acid sequence homology in their p40 and p35 subunits, respectively. IL-12 apparently shows species specificity with human IL-12 reportedly showing minimal activity in the murine system. IL-12 is produced by macrophages and B lymphocytes and has been shown to have multiple effects on T cells and natural killer (NK) cells (4). These effects include inducing production of IFN-gamma and TNF by resting and activated T and NK cells, synergizing with other IFN‑gamma inducers at both the transcriptional and post-transcriptional levels. This interaction induces IFN-gamma gene expression, enhancing the cytotoxic activity of resting NK and T cells, inducing and synergizing with IL-2 in the generation of lymphokine-activated killer (LAK) cells, acting as a co-mitogen to stimulate proliferation of resting T cells, and inducing proliferation of activated T and NK cells (5). Current evidence indicates that IL‑12, produced by macrophages in response to infectious agents, is a central mediator of the cell‑mediated immune response by its actions on the development, proliferation, and activities of TH1 cells. In its role as the initiator of cell-mediated immunity, it has been suggested that IL-12 has therapeutic potential as a stimulator of cell-mediated immune responses to microbial pathogens, metastatic cancers, and viral infections such as AIDS.

Item	Value
References	Gubler, U. et al. (1991) Proc. Natl. Acad. Sci. 88:4143. Gearing, D. et al. (1991) Cell 66:9. Merberg, D. et al. (1992) Immunology Today 13:78. Wolf, S.F. et al. (1991) Journal of Immunology 146:3074. Airoldi, I. et al. (2000) Journal of Immunology 165:6880.
Long Name	Interleukin 12
Entrez Gene IDs	3592 (Human); 16159 (Mouse); 84405 (Rat); 397053 (Porcine); 403977 (Canine); 102115316 (Cynomolgus Monkey); 493741 (Feline); 703205 (Rhesus Macaque)
Alternate Names	CLMF p35, CLMF1, Cytotoxic lymphocyte maturation factor 35 kDa subunit, IL-12, IL-12 subunit p35, IL-12, subunit p35, IL12, IL12A, NF cell stimulatory factor chain 1, NK cell stimulatory factor chain 1, NKSF1, interleukin 12, p35, interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35), interleukin-12 alpha chain, interleukin-12 subunit alpha, natural killer cell stimulatory factor 1, 35 kD subunit, p35

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FAQs

What is the difference between Recombinant Human IL-12 Protein (Catalog # 219-IL) and Recombinant Human Il-12 (linked heterodimer) Protein (Catalog # 10018-IL?

The Recombinant Human IL-12 Protein (Catalog # 219-IL), expressed in Sf 21 cells, is a disulfide-linked homodimer consisting of the p35 and p40 subunits. The subunits can be separated under reducing conditions. The Recombinant Human IL-12 (linked heterodimer) Protein (Catalog # 10018-IL), expressed in HEK293 cells, is a chimeric protein consisting of the p35 and p40 subunits connected by a linker. These two subunits cannot be separated.