Sandwich ELISA analysis of human IL-12 was performed using a Human IL-12 (p70) colorimetric ELISA kit (Product # EHIL12) by loading 50 ul per well of a biotinylated anti-human IL-12 antibody (Product # M121B) in duplicate at 0.03125 ug/ml, and 50 ul 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 ug/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 ul 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.
|Tested species reactivity||Human|
|Published species reactivity||Bacteria, Human|
|Host / Isotype||Rat / IgG1|
|Immunogen||Recombinant human IL-12 (p70)|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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 gene encodes a subunit of a cytokine that acts on T and natural killer cells, and has a broad array of biological activities. The cytokine 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. This cytokine is required for the T-cell-independent induction of interferon (IFN)-gamma, and is important for the differentiation of both Th1 and Th2 cells. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
IL-12 receptor ß1 deficiency alters in vivo T follicular helper cell response in humans.
M122 was used in blocking or activating experiment to study human T follicular helper cell responses in individuals lacking IL-Rbeta1
|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||Blood (121:3375)||2013|
Interleukin-12 production by leukemia-derived dendritic cells counteracts the inhibitory effect of leukemic microenvironment on T cells.
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
|Curti A,Pandolfi S,Aluigi M,Isidori A,Alessandrini I,Chiodoni C,Testoni N,Colombo MP,Baccarani M,Lemoli RM||Experimental hematology (33:1521)||2005|
Trypanosoma cruzi (Chagas' disease agent) reduces HIV-1 replication in human placenta.
M122 was used in ELISA to study the role of Trypanosoma cruzi in HIV-1 replication in human placenta
|Dolcini GL,Solana ME,Andreani G,Celentano AM,Parodi LM,Donato AM,Elissondo N,González Cappa SM,Giavedoni LD,Martínez Peralta L||Retrovirology (5:null)||2008|
Interleukin 6 and 12, alanine aminotransferase activity, and HCV viral load in children with chronic hepatitis C treated with interferon and ribavirin.
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
|Kowala-Piaskowska A,Mozer-Lisewska I,Figlerowicz M,Machowska L,S¿uzewski W||Inflammation (28:319)||2004|
High production of proinflammatory and Th1 cytokines by dendritic cells from patients with rheumatoid arthritis, and down regulation upon FcgammaR triggering.
M122 was used in ELISA to investigate the production of proinflammatory and Th1 cytokines by dendritic cells from patients with rheumatoid arthritis
|Radstake TR,van Lent PL,Pesman GJ,Blom AB,Sweep FG,Rönnelid J,Adema GJ,Barrera P,van den Berg WB||Annals of the rheumatic diseases (63:696)||2004|
Vitamin E-loaded dialyzer resets PBMC-operated cytokine network in dialysis patients.
M122 was used in ELISA to study the effect of vitamin E on cytokine profile in dialysis patients
|Libetta C,Zucchi M,Gori E,Sepe V,Galli F,Meloni F,Milanesi F,Dal Canton A||Kidney international (65:1473)||2004|
Monocyte chemoattractant protein-1 levels in bronchoalveolar lavage fluid of lung-transplanted patients treated with tacrolimus as rescue treatment for refractory acute rejection.
M122 was used in ELISA to investigate the effect of immunosuppressive regimens on cytokine expression in lung transplanted patients
|Meloni F,Cascina A,Paschetto E,Marone Bianco A,Morosini M,Pellegrini C,Fietta A,Vitulo P,Pozzi E,Viganò M||Transplantation proceedings (35:1523)||2003|
Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.
M122 was used in ELISA to investigate the augmented production of Th1-type cytokines in HCV-infected cryoglobulinemic patients
|Loffreda S,Muratori P,Muratori L,Mele L,Bianchi FB,Lenzi M||Journal of hepatology (38:230)||2003|
Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent.
M122 was used in ELISA to investigate the mechanism for the effect of Leishmania on interleukin 12 p70 secretion
|McDowell MA,Marovich M,Lira R,Braun M,Sacks D||Infection and immunity (70:3994)||2002|
Monocyte induction of IL-10 and down-regulation of IL-12 by iC3b deposited in ultraviolet-exposed human skin.
M122 was used in ELISA to investigate the regulation of interleukin-10 and interleukin-12 by iC3b deposited in ultraviolet-exposed human skin
|Yoshida Y,Kang K,Berger M,Chen G,Gilliam AC,Moser A,Wu L,Hammerberg C,Cooper KD||Journal of immunology (Baltimore, Md. : 1950) (161:5873)||1998|
IL-12-deficient dendritic cells, generated in the presence of prostaglandin E2, promote type 2 cytokine production in maturing human naive T helper cells.
M122 was used in ELISA to investigate the importance of PGE2 during dendritic cell development
|Kali¿ski P,Hilkens CM,Snijders A,Snijdewint FG,Kapsenberg ML||Journal of immunology (Baltimore, Md. : 1950) (159:28)||1997|
Reduced production of IL-12 and IL-12-dependent IFN-gamma release in patients with allergic asthma.
M122 was used in ELISA to investigate the expression level of interleukin 12 and interferon gamma in patients with allergic asthma
|van der Pouw Kraan TC,Boeije LC,de Groot ER,Stapel SO,Snijders A,Kapsenberg ML,van der Zee JS,Aarden LA||Journal of immunology (Baltimore, Md. : 1950) (158:5560)||1997|
Reduced IL-12 production by monocytes upon ultraviolet-B irradiation selectively limits activation of T helper-1 cells.
M122 was used in ELISA to investigate the influence of UVB irradiation on the activation of human T helper cells
|Kremer IB,Hilkens CM,Sylva-Steenland RM,Koomen CW,Kapsenberg ML,Bos JD,Teunissen MB||Journal of immunology (Baltimore, Md. : 1950) (157:1913)||1996|
Regulation of bioactive IL-12 production in lipopolysaccharide-stimulated human monocytes is determined by the expression of the p35 subunit.
M122 was used in ELISA to investigate the regulation of interleukin 12 expression
|Snijders A,Hilkens CM,van der Pouw Kraan TC,Engel M,Aarden LA,Kapsenberg ML||Journal of immunology (Baltimore, Md. : 1950) (156:1207)||1996|
Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells.
M122 was used in ELISA to investigate the expression of IL-12 by peripheral blood mononuclear cells
|D'Andrea A,Rengaraju M,Valiante NM,Chehimi J,Kubin M,Aste M,Chan SH,Kobayashi M,Young D,Nickbarg E||The Journal of experimental medicine (176:1387)||1992|