Invitrogen™ IL-12 p70 Human Uncoated ELISA Kit

Uncoated ELISA Kit

Catalog Number	Mfr. No.	Quantity
50-172-60	Invitrogen™ 88712688	10 x 96 Tests
50-172-58	Invitrogen™ 88712677	20 x 96 Tests

Description

The Human Interleukin 12 p70 (IL-12 p70) Uncoated ELISA Kit contains pre-matched antibody pairs, and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human IL-12 p70. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately. Principle of the method ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

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.

Specifications

• Accession Number: P29459,P29460
• Assay Sensitivity: 4 pg/mL
• Product Type: ELISA Kit
• Target Species: Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene Symbol: IFN-g,Ifg
• Label or Dye: HRP
• Sample Volume: Plasma, 100 μL; Serum, 100 μL; Supernatant, 100 μL
• Synonym: Interleukin-12, IL12, IL23
• Test Time: 1 hr. 45 min.

• Assay Range: 4-500 pg/mL
• Conjugate: Biotin
• Sample Type: Plasma, Serum, Supernatant
• For Use With (Application): ELISA
• Gene ID (Entrez): 3592, 3593
• Kit Contents: Capture Antibody: Pre-titrated; purified antibody, Detection Antibody: Pre-titrated; biotin-conjugated antibody, Standard: Recombinant cytokine for generating standard curve and calibrating samples, ELISA/ELISPOT Coating Buffer Powder: This uncoated ELISA kit may contain ELISA/ELISPOT Coating Buffer Powder (Reconstitute to 1L with dH20 and filter (0.22 µM)) or 10X PBS ELISA Coating Buffer (Dilute 1 part 10X Buffer into 9 parts dH20)., Assay Diluent: 5X concentrated, Detection enzyme: Pre-titrated Streptavidin-HRP, Substrate Solution: Tetramethylbenzidine (TMB) Substrate Solution, Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards, 96 Well Plate: Corning Costar 9018 (included with product Cat. Nos. ending in suffixes -22; -44; -76; -86)
• Regulatory Status: RUO
• Storage Requirements: 2°C to 8°C
• Target: Human
• Total Assay Time: 24 hr. 30 min.