OAS1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level 2',5'-Oligoadenylate Synthetase 1 (OAS1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level 2',5'-Oligoadenylate Synthetase 1 (OAS1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

(2-5') oligo (A) synthase 1;2',5'-oligo A synthetase 1;2-5A synthase 1;2-5A synthetase 1;2'-5'-oligoadenylate synthase 1;2'-5'-oligoadenylate synthetase 1, 40/46kDa;2'-5'-oligoisoadenylate synthetase 1; E16 (2'-5') oligo A synthetase; E18/E16; IFI-4; OIAS; OIASI; p46/p42 OAS.

Interferon-induced, dsRNA-activated antiviral enzyme which plays a critical role in cellular innate antiviral response (PubMed:34581622) . In addition, it may also play a role in other cellular processes such as apoptosis, cell growth, differentiation and gene regulation. Synthesizes higher oligomers of 2'-5'-oligoadenylates (2-5A) from ATP which then bind to the inactive monomeric form of ribonuclease L (RNase L) leading to its dimerization and subsequent activation. Activation of RNase L leads to degradation of cellular as well as viral RNA, resulting in the inhibition of protein synthesis, thus terminating viral replication (PubMed:34581622) . Can mediate the antiviral effect via the classical RNase L-dependent pathway or an alternative antiviral pathway independent of RNase L. The secreted form displays antiviral effect against vesicular stomatitis virus (VSV), herpes simplex virus type 2 (HSV-2), and encephalomyocarditis virus (EMCV) and stimulates the alternative antiviral pathway independent of RNase L.

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 2',5'-Oligoadenylate Synthetase 1 (OAS1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to 2',5'-Oligoadenylate Synthetase 1 (OAS1) . 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 2',5'-Oligoadenylate Synthetase 1 (OAS1), 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 2',5'-Oligoadenylate Synthetase 1 (OAS1) 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) 92-99 95 EDTA Plasma (n=5) 78-92 86 Heparin Plasma (n=5) 92-101 96 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 92-99 95 Serum (n=5) 92-99 95 EDTA Plasma (n=5) 78-92 86 EDTA Plasma (n=5) 78-92 86 Heparin Plasma (n=5) 92-101 96 Heparin Plasma (n=5) 92-101 96

Component Amount Anti-OAS1 Microplate 96 Wells (12 x 8 Well Strips) OAS1 Lyophilized Standard 2 x 300 ng 100X Biotinylated OAS1 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-OAS1 Microplate 96 Wells (12 x 8 Well Strips) Anti-OAS1 Microplate 96 Wells (12 x 8 Well Strips) OAS1 Lyophilized Standard 2 x 300 ng OAS1 Lyophilized Standard 2 x 300 ng 100X Biotinylated OAS1 Detector Antibody 120 uL 100X Biotinylated OAS1 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 2',5'-Oligoadenylate Synthetase 1 (OAS1) . No significant cross-reactivity or interference between 2',5'-Oligoadenylate Synthetase 1 (OAS1) and analogues was observed.