Invitrogen

IL-12 p70 Monoclonal Antibody (20C2)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

2 Published Figures

20 References

View all (20) IL-12 p70 antibodies

Datasheet

Protocols

Questions & Answers

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Datasheet

Protocols

Questions & Answers

Tech Support

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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.

IL-12 p70 Antibody — Figure 2 TLR ligation induces differential cytokine production by human blood DCs. The DCs were stimulated as indicated and cultured overnight at 37degC. IL-10 and IL-12p70 production was analyzed in supernatants of overnight cultures by standard sandwich ELISA ( n >= 6). Each symbol represents one donor (also across the subsets). Significance was determined by Kruskal-Wallis test followed by Dunn's testing comparing different conditions of the same subset ( ** P < .01; *** P < .001). {TM}

Product Details

M122

Applications	Tested Dilution	Publications
Western Blot (WB)	Assay-dependent	-
ELISA (ELISA)	Assay-dependent	View 18 publications 18 publications
Neutralization (Neu)	-	View 2 publications 2 publications

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.

Product References

Human blood myeloid and plasmacytoid dendritic cells cross activate each other and synergize in inducing NK cell cytotoxicity.

Oncoimmunology

van Beek JJ,Gorris MA,Sköld AE,Hatipoglu I,Van Acker HH,Smits EL,de Vries IJ,Bakdash G

Published figure using IL-12 p70 monoclonal antibody (Product # M122) in ELISA

Tue Jan 12 00:00:00 EST 2021

Type 1-programmed dendritic cells drive antigen-specific latency reversal and immune elimination of persistent HIV-1.

EBioMedicine

Kristoff J,Palma ML,Garcia-Bates TM,Shen C,Sluis-Cremer N,Gupta P,Rinaldo CR,Mailliard RB

M122 was used in an ELISA assay to test the use of antigen-presenting type 1-polarized, monocyte-derived dendritic cells generated from chronic HIV-1-infected individuals on ART as a means to induce HIV-1 latency reversal in autologous CD4+ T cells harboring replication-competent provirus.

Wed May 01 00:00:00 EDT 2019

The Phosphoinositide Kinase PIKfyve Promotes Cathepsin-S-Mediated Major Histocompatibility Complex Class II Antigen Presentation.

iScience

Baranov MV,Bianchi F,Schirmacher A,van Aart MAC,Maassen S,Muntjewerff EM,Dingjan I,Ter Beest M,Verdoes M,Keyser SGL,Bertozzi CR,Diederichsen U,van den Bogaart G

M122 was used in an ELISA assay to study the role of PIKfyve in the processing and presentation of antigens.

Fri Jan 25 00:00:00 EST 2019

A Comparative Study of the T Cell Stimulatory and Polarizing Capacity of Human Primary Blood Dendritic Cell Subsets.

Mediators of inflammation

Sittig SP,Bakdash G,Weiden J,Sköld AE,Tel J,Figdor CG,de Vries IJ,Schreibelt G

Published figure using IL-12 p70 monoclonal antibody (Product # M122) in ELISA

Tue Jan 17 00:00:00 EST 2017

IL-12 receptor β1 deficiency alters in vivo T follicular helper cell response in humans.

Blood

Schmitt N,Bustamante J,Bourdery L,Bentebibel SE,Boisson-Dupuis S,Hamlin F,Tran MV,Blankenship D,Pascual V,Savino DA,Banchereau J,Casanova JL,Ueno H

M122 was used in blocking or activating experiment to study human T follicular helper cell responses in individuals lacking IL-Rbeta1

Thu Apr 25 00:00:00 EDT 2013

Trypanosoma cruzi (Chagas' disease agent) reduces HIV-1 replication in human placenta.

Retrovirology

Dolcini GL,Solana ME,Andreani G,Celentano AM,Parodi LM,Donato AM,Elissondo N,González Cappa SM,Giavedoni LD,Martínez Peralta L

M122 was used in ELISA to study the role of Trypanosoma cruzi in HIV-1 replication in human placenta

Tue Jul 01 00:00:00 EDT 2008

Interleukin-12 production by leukemia-derived dendritic cells counteracts the inhibitory effect of leukemic microenvironment on T cells.

Experimental hematology

Curti A,Pandolfi S,Aluigi M,Isidori A,Alessandrini I,Chiodoni C,Testoni N,Colombo MP,Baccarani M,Lemoli RM

M122 was used in blocking/activating experiment to investigate the role of interleukin-12 produced by leukemia-derived dendritic cells for the inhibitory effect of leukemic microenvironment on T cells

Thu Dec 01 00:00:00 EST 2005

Interleukin 6 and 12, alanine aminotransferase activity, and HCV viral load in children with chronic hepatitis C treated with interferon and ribavirin.

Inflammation

Kowala-Piaskowska A,Mozer-Lisewska I,Figlerowicz M,Machowska L,Słuzewski W

M122 was used in ELISA to study the effect of interferon and ribavirin treatment on interluekin 6 and 12, alanine aminotransferase activity, and HCV viral load

Wed Dec 01 00:00:00 EST 2004

High production of proinflammatory and Th1 cytokines by dendritic cells from patients with rheumatoid arthritis, and down regulation upon FcgammaR triggering.

Annals of the rheumatic diseases

Radstake TR,van Lent PL,Pesman GJ,Blom AB,Sweep FG,Rönnelid J,Adema GJ,Barrera P,van den Berg WB

M122 was used in ELISA to investigate the production of proinflammatory and Th1 cytokines by dendritic cells from patients with rheumatoid arthritis

Tue Jun 01 00:00:00 EDT 2004

Vitamin E-loaded dialyzer resets PBMC-operated cytokine network in dialysis patients.

Kidney international

Libetta C,Zucchi M,Gori E,Sepe V,Galli F,Meloni F,Milanesi F,Dal Canton A

M122 was used in ELISA to study the effect of vitamin E on cytokine profile in dialysis patients

Thu Apr 01 00:00:00 EST 2004

Altered maturation of peripheral blood dendritic cells in patients with breast cancer.

British journal of cancer

Della Bella S,Gennaro M,Vaccari M,Ferraris C,Nicola S,Riva A,Clerici M,Greco M,Villa ML

M122 was used in an ELISA assay to evaluate the in vivo relevance of two pathways in breast cancer patients.

Mon Oct 20 00:00:00 EDT 2003

Monocyte chemoattractant protein-1 levels in bronchoalveolar lavage fluid of lung-transplanted patients treated with tacrolimus as rescue treatment for refractory acute rejection.

Transplantation proceedings

Meloni F,Cascina A,Paschetto E,Marone Bianco A,Morosini M,Pellegrini C,Fietta A,Vitulo P,Pozzi E,Viganò M

M122 was used in ELISA to investigate the effect of immunosuppressive regimens on cytokine expression in lung transplanted patients

Sun Jun 01 00:00:00 EDT 2003

Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.

Journal of hepatology

Loffreda S,Muratori P,Muratori L,Mele L,Bianchi FB,Lenzi M

M122 was used in ELISA to investigate the augmented production of Th1-type cytokines in HCV-infected cryoglobulinemic patients

Sat Feb 01 00:00:00 EST 2003

Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent.

Infection and immunity

McDowell MA,Marovich M,Lira R,Braun M,Sacks D

M122 was used in ELISA to investigate the mechanism for the effect of Leishmania on interleukin 12 p70 secretion

Thu Aug 01 00:00:00 EDT 2002

Monocyte induction of IL-10 and down-regulation of IL-12 by iC3b deposited in ultraviolet-exposed human skin.