Mouse Isoleucyl tRNA Synthetase ELISA Kit

From Assay Genie, this Sandwich ELISA (Double Antibody) kit is designed for the quantitative measurement of mouse Isoleucyl tRNA Synthetase in mouse samples. Validated sample types include serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Detection is colorimetric, using a sandwich ELISA format in which a biotinylated detection antibody and HRP-conjugated Streptavidin are combined with TMB substrate and read at 450 nm.

Reported performance figures include a sensitivity of 0.375 ng/mL and a working range of 0.625-40 ng/mL. Linearity across 1:2, 1:4 and 1:8 dilutions for serum (n=5) was 88-105%, 90-104% and 89-104%; for EDTA plasma (n=5) was 90-101%, 82-97% and 89-101%; and for heparin plasma (n=5) was 87-100%, 82-87% and 81-97%. Recovery: serum (n=5) ranged 89-102% with an average of 96%; EDTA plasma (n=5) ranged 85-98% with an average of 91%; heparin plasma (n=5) ranged 85-98% with an average of 91%. The kit is stored at 2-8°C for 12 months. Associated Uniprot reference: Q8BU30. Research area: Metabolism. Synonyms include IARS, Isoleucine--tRNA ligase, IRS, IleRS, and Isoleucyl-tRNA synthetase, cytoplasmic.

The assay workflow is: equilibrate reagents and TMB substrate to room temperature and set up standards, samples and zero controls on the pre-coated plate; incubate at 37°C for 90 minutes for antigen binding; add biotin-labeled detection antibody and incubate at 37°C for 60 minutes; add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes; develop with TMB substrate in the dark for 10–20 minutes; stop the reaction and read absorbance at 450 nm. Sample preparation guidance covers serum (clot, centrifuge at 1000 × g for 20 minutes), plasma (anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate, cell culture supernatant (centrifuge at 2500 rpm for 5 minutes) and cell lysate workflows.