IL12 antibody

Catalog No. ABIN2689731

Product Details anti-IL12 Antibody

Target: IL12 (Interleukin 12 (IL12))

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This IL12 antibody is un-conjugated

Application: Blocking Antibody (Inhibition)

Brand: BD Pharmingen™

Characteristics: The C15.6 antibody reacts with both free and complexed (homodimer p80 and heterodimer p70) forms of the p40 subunit of mouse interleukin-12 (IL-12). The immunogen used to generate the C15.6 hybridoma was recombinant mouse IL-12 p70 protein. p40 has also been described as a subunit of IL-23 and thus it is possible that the C15.6 antibody will crossreact with IL-23. This antibody is routinely tested by flow cytometric analysis. Other applications were tested during antibody development only or reported in the literature. Expression of IL-12 by mouse bone marrow-derived macrophages. Bone marrow cells from 6 month old BALB/c mice were cultured for 10 days in mouse GM-CSF (40 ng/mL final concentration, Cat. No. 554586). Adherent cells were washed and treated for ~14 hours with mouse IFN-γ (10 ng/mL final concentration, Cat. No. 554587), subsequently LPS (1 μg/mL final concentration, Sigma) and GolgiStop™ (2 μM final concentration Cat. No. 554724) were added to cultures for an additional 5 hours. Adherent cells were washed and then incubated with 1x trypsin EDTA at 37 °C. for 15 minutes and gently dislodged by pipetting. Nonspecific surface binding was blocked by incubation of cells with purified polyclonal normal mouse immunoglobulin. Cells were then fixed, permeabilized, and non-specific binding to intracellular antigens was blocked using PBS/2 % BSA/0.1 % saponin. Cells were then stained with 0.06 μg of PE-conjugated rat anti-mouse IL-12 (p40/p70) antibody (PE-C15.6, Cat. No. 554479, see left panel) using the BD Pharmingen™ staining protocol. To demonstrate specificity of staining, the binding of PE-C15.6 antibody was blocked by the preincubation of the conjugated antibody with recombinant mouse IL-12 p40 protein (0.5 μg, Cat. No. 554594, see middle panel), and by preincubating the fixed/permeabilized cells with unlabeled C15.6 antibody (5 μg/mL final concentration, Cat. No. 554477, see right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the ligand blocking and unlabeled antibody blocking controls.

BD Pharmingen™ Purified Rat Anti-Mouse IL-12 (p40/p70) - Purified - Clone C15.6 - Isotype Rat IgG1 - Reactivity Ms - 0.1 mg

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: CHO-expressed recombinant mouse IL-12 p70 protein

Clone: C15-6

Isotype: IgG1

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Handling

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C.

References for anti-IL12 antibody (ABIN2689731)

Prussin, Metcalfe: "Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies." in: Journal of immunological methods, Vol. 188, Issue 1, pp. 117-28, (1996) (PubMed).

Wysocka, Kubin, Vieira, Ozmen, Garotta, Scott, Trinchieri: "Interleukin-12 is required for interferon-gamma production and lethality in lipopolysaccharide-induced shock in mice." in: European journal of immunology, Vol. 25, Issue 3, pp. 672-6, (1995) (PubMed).

Target Details for IL12

Target: IL12 (Interleukin 12 (IL12))

Alternative Name: IL-12 p40,p70 (IL12 Products)

Synonyms: CLMF antibody, CLMF2 antibody, IL-12B antibody, NKSF antibody, NKSF2 antibody, p40 antibody, Il-12b antibody, Il12p40 antibody, Il-12p40 antibody, Il12 antibody, IL-12p40 antibody, IL-12 antibody, LOC100217488 antibody, IL-12p35 antibody, Il-12a antibody, Ll12a antibody, p35 antibody, IL12p40 antibody, IL12B antibody, IL-12 p35 antibody, interleukin 12B antibody, interleukin 12A antibody, interleukin 12b antibody, interleukin 12a antibody, Interleukin 12 level antibody, IL12B antibody, IL12A antibody, Il12b antibody, Il12a antibody, IL12 antibody, il12a antibody

Pathways: JAK-STAT Signaling, TLR Signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Activated T Cell Proliferation, Cancer Immune Checkpoints, Inflammasome