Invitrogen

IL-12 p70 Monoclonal Antibody (20C2)

Product Image Gallery

IL-12 p70 Antibody — Figure 1. Human CD1c + DCs and pDCs cross activate each other. CD1c + DCs and pDCs were cultured overnight, either separately or together, in the presence of indicated stimuli. Maturation was checked on the specific DC subsets within the co-culture. (A) Relative expression of CD86, CD80, PD-L1, and MHC class I and II on pDCs in geometric MFI normalized to pDCs cultured alone with IL-3. (B) Relative expression of CD86, CD80, PD-L1 and MHC class I and II on CD1c + DCs in geometric MFI normalized to CD1c + DCs cultured alone with GM-CSF. (C) IL-12p70, IFN-alpha, and TNF-alpha in supernatants of overnight cultures were analyzed by ELISA. Black bars, pDCs; white bars, CD1c + DCs; gray bars, CD1c + DCs and pDCs. Results are the mean +- SEM of at least five (A, B) or at least three (C) independent experiments. Significance was determined by repeated measures one-way ANOVA, followed by a post-hoc Dunnett's test (* p < 0.05; ** p < 0.01; *** p < 0.001). Only conditions that are significantly different when compared with extra control groups (see Fig. S1) are denoted in the graphs.

IL-12 p70 Antibody in ELISA (ELISA) — Sandwich ELISA analysis of human IL-12 was performed using a Human IL-12 (p70) colorimetric ELISA kit (Product # EHIL12) by loading 50 µL per well of a biotinylated anti-human IL-12 antibody (Product # M121B) in duplicate at 0.03125 µg/mL, and 50 µL per well of Human IL-12 (p70) Recombinant Protein (Product # SIL12) in duplicate at 600, 240, 96, 38.4, 15.4, 0 pg/mL across a 7 µg/mL anti-human IL-12 (Product # M122) pre-coated plate and incubating for 3 hours at room temperature. The plate was washed then incubated with 100 µL per well of Streptavidin-HRP (Product # N504) in all test wells at 1:3,000 for 30 minutes at room temperature. Detection was performed with TMB substrate for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Product Details

Tested Applications

Dilution

ELISA (ELISA)

Assay Dependent

Western Blot (WB)

Assay Dependent

Published Applications

ELISA (ELISA)

See 15 publications below

Neutralization (Neu)

See 2 publications below

Product Specifications

Species Reactivity

Human

Published species

Bacteria, Human

Host / Isotype

Rat / IgG1

Class

Monoclonal

Type

Antibody

Clone

20C2

Immunogen

Recombinant human IL-12 (p70)

Conjugate

Unconjugated

Form

Liquid

Concentration

1.0 mg/mL

Purification

Protein G

Storage buffer

PBS

Contains

no preservative

Storage conditions

-20°C

RRID

AB_223556

Product Specific Information

M122 targets IL-12 in ELISA, and WB applications and shows reactivity with Human samples.

The M122 immunogen is recombinant human IL-12 (p70).

M122 detects IL-12 which has a predicted molecular weight of approximately 23 kDa.

The M122 IL12 antibody (clone 20C2) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M121B (biotinylated conjugate of clone C8.6) Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection.

Antibody M122 (clone 20C2) and biotinylated antibody M121B (clone C8.6) have successfully been used in combination with recombinant IL12 p70 protein SIL12 in ELISA applications.

This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).

Target Information

Interleukin-12 (IL-12) is a heterodimeric 70 kDa cytokine composed of two covalently linked, glycosylated chains with molecular weights of 40kD (p40) and 35-kD (p35). IL-12 is mainly produced by monocytes, macrophages, and dendritic cells in response to bacterial products such as lipopolysaccharides (LPS), to intracellular pathogens or upon interaction with activated T cells. IL-12 was originally discovered because of its ability to induce interferon-gamma (IFN-g) production, cell proliferation, and cytotoxicity mediated by natural killer cells and T cells. Studies have established that IL-12 also plays a key role in the development of Th1 responses, leading to IFN-g and IL-2 production. These cytokines can in turn promote T-cell responses and macrophage activation. Recombinant mouse IL-12 p70 is produced in baculovirus-infected insect cells as an authentic heterodimer of precursor p35 and p40 subunits using a dual promoter expression system. IL-12 p70 is distinct from other available forms of the protein in that it is expressed as a true heterodimer, as opposed to a single-chain, pseudo-heterodimer in which the subunits are joined by an artificial linker. The responses of lymphocytes to this cytokine are mediated by the activator of transcription protein STAT4. Nitric oxide synthase 2A (NOS2A/NOS2) is found to be required for the signaling process of this cytokine in innate immunity. Human and mouse IL-12 p40 share about 70% amino acid sequence homology.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: 20C2; CLMF p35; CLMF p40; cytotoxic lymphocyte maturation factor 1, p35; Cytotoxic lymphocyte maturation factor 35 kDa subunit; Cytotoxic lymphocyte maturation factor 40 kDa subunit; IL-12 subunit p35; IL-12 subunit p40; IL-12, subunit p35; IL-12A; IL-12B; IL12, subunit p40; IL12p35; IL12P40; IL35 subunit; interleukin 12, p35; interleukin 12, p40; interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocyte maturation factor 1, p35); interleukin 12B (natural killer cell stimulatory factor 2, cytotoxic lymphocyte maturation factor 2, p40); interleukin-12 alpha chain; interleukin-12 beta chain; Interleukin-12 subunit alpha; Interleukin-12 subunit beta; natural killer cell stimulatory factor 1, 35 kD subunit; natural killer cell stimulatory factor, 40 kD subunit; NF cell stimulatory factor chain 1; NK cell stimulatory factor chain 1; NK cell stimulatory factor chain 2; NKSF1; NKSF2

Gene Aliases: CLMF; CLMF2; IL-12A; IL-12B; IL12A; IL12B; IMD28; IMD29; NFSK; NKSF; NKSF1; NKSF2; P35

UniProt ID: (Human) P29459, (Human) P29460

Entrez Gene ID: (Human) 3592, (Human) 3593

Molecular Function: cytokine interleukin superfamily signaling molecule