GARS ELISA Kit (Human)

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

Ap4A synthetase; AP-4-A synthetase; Charcot-Marie-Tooth neuropathy 2D; Charcot-Marie-Tooth neuropathy, neuronal type, D; CMT2D; diadenosine tetraphosphate synthetase; DSMAV; GARS; glycine--tRNA ligase; Glycyl-tRNA synthetase; Glycyl-tRNA synthetase 1; GlyRS; HMN5; HMN5A; SMAD1; SMAJI.

Catalyzes the ATP-dependent ligation of glycine to the 3'-end of its cognate tRNA, via the formation of an aminoacyl-adenylate intermediate (Gly-AMP) (PubMed:17544401, PubMed:28675565, PubMed:24898252) . Also produces diadenosine tetraphosphate (Ap4A), a universal pleiotropic signaling molecule needed for cell regulation pathways, by direct condensation of 2 ATPs. Thereby, may play a special role in Ap4A homeostasis (PubMed:19710017) .

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 Glycyl tRNA Synthetase (GARS) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glycyl tRNA Synthetase (GARS) . 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 Glycyl tRNA Synthetase (GARS), 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 Glycyl tRNA Synthetase (GARS) 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) 80-98 94 EDTA Plasma (n=5) 94-103 98 Heparin Plasma (n=5) 90-99 93 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 80-98 94 Serum (n=5) 80-98 94 EDTA Plasma (n=5) 94-103 98 EDTA Plasma (n=5) 94-103 98 Heparin Plasma (n=5) 90-99 93 Heparin Plasma (n=5) 90-99 93

Component Amount Anti-GARS Microplate 96 Wells (12 x 8 Well Strips) GARS Lyophilized Standard 2 x 1 ng 100X Biotinylated GARS 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-GARS Microplate 96 Wells (12 x 8 Well Strips) Anti-GARS Microplate 96 Wells (12 x 8 Well Strips) GARS Lyophilized Standard 2 x 1 ng GARS Lyophilized Standard 2 x 1 ng 100X Biotinylated GARS Detector Antibody 120 uL 100X Biotinylated GARS 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 Glycyl tRNA Synthetase (GARS) . No significant cross-reactivity or interference between Glycyl tRNA Synthetase (GARS) and analogues was observed.