IL1RN ELISA Kit (Dog)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Interleukin 1 Receptor Antagonist (IL1RA) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Interleukin 1 Receptor Antagonist (IL1RA) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Inhibits the activity of interleukin-1 by binding to receptor IL1R1 and preventing its association with the coreceptor IL1RAP for signaling. Has no interleukin-1 like activity (By similarity) .

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 1 Receptor Antagonist (IL1RA) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Interleukin 1 Receptor Antagonist (IL1RA) . 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 1 Receptor Antagonist (IL1RA), 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 1 Receptor Antagonist (IL1RA) 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) 83-99 93 EDTA Plasma (n=5) 89-96 92 Heparin Plasma (n=5) 88-99 94 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 83-99 93 Serum (n=5) 83-99 93 EDTA Plasma (n=5) 89-96 92 EDTA Plasma (n=5) 89-96 92 Heparin Plasma (n=5) 88-99 94 Heparin Plasma (n=5) 88-99 94

Component Amount Anti-IL1RN Microplate 96 Wells (12 x 8 Well strips) IL1RN Lyophilized Standard 2 100X Biotinylated IL1RN Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-IL1RN Microplate 96 Wells (12 x 8 Well strips) Anti-IL1RN Microplate 96 Wells (12 x 8 Well strips) IL1RN Lyophilized Standard 2 IL1RN Lyophilized Standard 2 100X Biotinylated IL1RN Detector Antibody 1 x 120 uL 100X Biotinylated IL1RN Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Interleukin 1 Receptor Antagonist (IL1RA) . No significant cross-reactivity or interference between Interleukin 1 Receptor Antagonist (IL1RA) and analogues was observed.