Invitrogen™ Human IL-27 ELISA Kit

Description

Includes

Aluminium pouch(es) with a Microwell Plate coated with monoclonal antibody to human IL-27
Biotin-Conjugate anti-human IL-27 monoclonal antibody
Streptavidin-HRP
Human IL-27 Standard lyophilized, 4ng/mL upon reconstitution
Sample Diluent
Calibrator Diluent
Assay Buffer Concentrate 20x(PBS with 1% Tween 20 and 10% BSA)
Wash Buffer Concentrate 20x(PBS with 1% Tween 20)
Substrate Solution (tetramethyl-benzidine)
Stop Solution (1M Phosphoric acid)
Blue-Dye
Green-Dye
Red-Dye
Adhesive Films

ELISA

The Human Interleukin-27 (Hu IL-27) ELISA quantitates Hu IL-27 in human serum, plasma, buffered solution, or cell culture medium. The assay will exclusively recognize both natural and recombinant Hu IL-27. Principle of the method The Human IL-27 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The 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. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

IL-27, a member of the IL-12 family, is a heterodimeric protein consisting of the p40-related protein Epstein-Barr virus-induced gene 3 (EBI3) non-covalently linked to an IL-12p35-related protein, p28 (also known as IL-30). IL-27 is produced by activated APCs and mature dendritic cells. IL-27 exerts its activities on NK cells and naive CD4+ T cells; mRNA expression analysis of IL-27 receptor components (WSX-1/TCCR and gp130) suggests that IL-27 may also target other cells, including mast cells and monocytes. Binding of IL-27 to WSX-1/gp130 activates JAK1, STAT1, and STAT3 and STAT1/3 phosphorylation. WSX-1/TCCR-deficient mice develop impaired Th1 responses and are more susceptible to infection with L. monocytogenes suggesting that Th1 responses require IL-27. Although activation of WSX-1 is required for the initiation of Th1 responses, it is not necessary for maintaining Th1 responses. IL-27 alone is not able to induce the differentiation of CD4+ T cells into IFN-γ-producing cells, suggesting a role for IL-27 as an initial activator of Th1 responses. An important effect of IL-27 in initiating Th1 responses is the induction of the Th1-specific transcription factor T-bet as well as the suppression of the Th2-specific transcription factor GATA-3. T-bet plays a critical role in Th1 differentiation by its ability to maintain IL-12Rβ2 expression following CD4+ T cell activation. Recent studies indicate that IL-27 has a potent antitumor activity. In vitro, IL-27 has been found to act directly on naive CD8 cells, generating CTL with enhanced granzyme B expression. In vivo, IL-27 has been reported to augment CTL activity, inhibit tumor growth, and induce complete regression of primary and metastatic neuroblastoma tumors.

Specifications

• Accession Number: Q14213,Q8NEV9
• Assay Range: 62.5 to 4,000 pg/mL
• Assay Sensitivity: 9.5 pg/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Plasma, Serum, Supernatant
• Target Species: Human
• For Use With (Application): ELISA
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 246778
• Gene Symbol: IL-27 B,IL27B
• Interassay CV: 5.6%
• Intraassay CV: 8.5
• Kit Contents: Pre-coated 96 well plate, Standard, Sample Diluent, Assay Buffer concentrate, Biotinylated Detection Antibody, SAV-HRP, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Plasma 50 μL, Serum 50 μL, Supernatant 50 μL
• Storage Requirements: 2°C to 8°C
• Synonym: Interleukin-27, IL27p28
• Target: IL-27 p28
• Test Time: 1 hrs 20 mins
• Total Assay Time: 3 hrs 30 mins

Safety and Handling

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