Sandwich ELISA analysis of Human IL-12 Biotinylated Antibody (Product # M121B) was performed using the Thermo Scientific Human IL-12 (p70) ELISA Kit (Product # EHIL12) by loading 50µl per well of Human IL-12 Biotinylated Antibody (Product # M121B) in duplicate at 0.03125µg/ml dilution and 50µl per well of Human IL-12 (p70) ELISA Standard (Product # SIL12) in duplicate at 600, 240, 96, 38.4, 15.4, 0 pg/ml across a pre-coated plate of Human IL-12 MAb (Product # M122) at 7µg/ml dilution and incubated for 3 hours at room temperature. The plate was washed then incubated with 100µl per well of Streptavidin-HRP (Product # N504) in all test wells at 1:3,000 dilution for 30 minutes at room temperature followed by 100µl per well of TMB substrate for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rat / IgG1|
|Immunogen||Recombinant human IL-12 (p70)|
|Storage buffer||PBS with 4% BSA|
|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.
|ELISA (ELISA)||See 8 publications below|
M121B targets IL-12 in ELISA, and WB applications and shows reactivity with Human samples.
The M121B immunogen is recombinant human IL-12 (p70).
M121B detects IL-12 which has a predicted molecular weight of approximately 23 kDa.
The M121B IL12 antibody (biotinylated conjugate of clone C8.6) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M122 (clone 20C2). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Biotinylated antibody M121B (clone C8.6) and antibody M122 (clone 20C2) have successfully been used in combination with recombinant IL12 p70 protein SIL12 in ELISA applications.
This antibody is produced by injecting Rat IgG secreting hybridoma cells into the peritoneum of mice. The resulting ascites is collected from the mice and the antibody is purified.
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.
Interleukin 6 and 12, alanine aminotransferase activity, and HCV viral load in children with chronic hepatitis C treated with interferon and ribavirin.
M121B 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.
M121B 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|
Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent.
M121B 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.
M121B 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.
M121B 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 IL-12 production by monocytes upon ultraviolet-B irradiation selectively limits activation of T helper-1 cells.
M121B 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.
M121B 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|
Natural killer cell stimulatory factor (interleukin 12 [IL-12]) induces T helper type 1 (Th1)-specific immune responses and inhibits the development of IL-4-producing Th cells.
M121B was used in ELISA to investigate the role of interleukin 12 on immune responses and Th cell development
|Manetti R,Parronchi P,Giudizi MG,Piccinni MP,Maggi E,Trinchieri G,Romagnani S||The Journal of experimental medicine (177:1199)||1993|