IL18 ELISA Kit (Horse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Interleukin 18 (IL18) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Interleukin 18 (IL18) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

IFN-gamma-inducing factor; IGIF; IL-1 gamma; IL-18; interferon gamma-inducing factor; interleukin 18 (interferon-gamma-inducing factor) ; interleukin-1 gamma; interleukin-18.

A proinflammatory cytokine primarily involved in polarized T-helper 1 (Th1) cell and natural killer (NK) cell immune responses. Upon binding to IL18R1 and IL18RAP, forms a signaling ternary complex which activates NF-kappa-B, triggering synthesis of inflammatory mediators. Synergizes with IL12/interleukin-12 to induce IFNG synthesis from T-helper 1 (Th1) cells and natural killer (NK) cells.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Interleukin 18 (IL18) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Interleukin 18 (IL18) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Interleukin 18 (IL18), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Interleukin 18 (IL18) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) Serum (n=5) 99-105 102 EDTA Plasma (n=5) 91-103 95 Heparin Plasma (n=5) 81-94 84 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 99-105 102 Serum (n=5) 99-105 102 EDTA Plasma (n=5) 91-103 95 EDTA Plasma (n=5) 91-103 95 Heparin Plasma (n=5) 81-94 84 Heparin Plasma (n=5) 81-94 84

Component Amount Anti-IL18 Microplate 96 Wells (12 x 8 Well Strips) IL18 Lyophilized Standard 2 x 1 ng 100X Biotinylated IL18 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-IL18 Microplate 96 Wells (12 x 8 Well Strips) Anti-IL18 Microplate 96 Wells (12 x 8 Well Strips) IL18 Lyophilized Standard 2 x 1 ng IL18 Lyophilized Standard 2 x 1 ng 100X Biotinylated IL18 Detector Antibody 120 uL 100X Biotinylated IL18 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Interleukin 18 (IL18) . No significant cross-reactivity or interference between Interleukin 18 (IL18) and analogues was observed.