Information Serum is lipid extracted to improve clarity, dialyzed against phosphate buffered saline (PBS), and freeze dried. Serum was collected before August 2019, prior to SARS-CoV-2 pandemic, and tested by ELISA for antibodies to SARS-CoV-2 Spike S1 Receptor Binding Domain (RBD).Usage Physical State: Freeze-dried solid

Storage and Rehydration: Store freeze-dried solid at 2-8°C. Rehydrate with the indicated volume of dH2O (see product specification sheet) and centrifuge if not clear. Prepare working dilution on day of use. Aliquot and freeze undiluted product at -20°C or below. Avoid repeated freezing and thawing.Expiration date: one year from date of rehydration. The expiration date may be extended if test results are acceptable for the intended use.

Buffer: 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6 Preservative:None. Product is not sterile, but may be sterile-filtered or a preservative may be added. Suggested Working Concentration or Dilution Range:1:100 - 1:400 for SARS-CoV-2 antibody ELISAs. Dilution should be optimized for assay of interest. Rehydration according to instruction yields 100% serum.Images & References

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* Apparent antibody concentration quantified compared to a standard curve generated with SARS-CoV-2 Spike S1 Antibody (HC2001) – Genscript A02038-100 Figure legend: ELISA for SARS-CoV-2 antibodies from human serum was carried out as described in Amanat et al. (2020) with the following modifications:Coating/capture material: SARS-CoV-2 Spike Protein RBD (Genscript – Z03483-100) diluted to 2 µg/ml in PBS, and 50 µl (2 µg/well) was coated onto an ELISA plate overnight at 4˚C. Plates were blocked with 1% Bovine Serum Albumin (IgG-Free, Protease-Free) (001-000-162) in PBS + 0.05% Tween-20 (PBST). Samples: Human IgG sourced pre-August 2019 [ChromPure Human IgG, whole molecule (009-000-003)] was diluted to 250 µg/ml in 1% BSA/PBST, in triplicate, (100 µl/well). Human Serum sourced pre-August 2019 (009-000-243) was diluted 1:150 in 1% BSA/PBST in triplicate (100 µl/well). Human serum confirmed to be SARS-CoV-2 naive was drawn from seven volunteers. Human serum deemed to be SARS-CoV-2 antibody positive (~ 26-37 days after exposure to SARS-CoV-2, confirmed diagnosis) was drawn from four individuals. Volunteer-derived human serum samples were diluted 1:150 in 1% BSA/PBST (100 µl/well).Detection: HRP-Conjugated secondary antibody (109-035-064) was diluted in PBST at 1:20,000 and added to each well (100 µl/well). Plates were incubated with TMB (Moss TMBHK-1000) for 30 minutes at room temperature. TMB was detected at 620 nm using a plate reader.Standard/calibration curve:Standard curves were generated using Human IgG SARS-CoV-2 Spike S1 Antibody (HC2001) (Genscript A02038). The antibody was serially diluted (1:3) in 1% BSA/PBST, in triplicate, across the ELISA plate starting at 200 ng/ml (100 µl/well).Analysis: Graphpad Prism 8 was used to fit non-linear regression curves to a standard curve generated using the anti-SARS-CoV-2 antibody. Results and apparent antibody concentrations (µg/ml) in serum were interpolated from the curves generated.

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This product is for in vitro research use only. It is not a medical device and it is not intended for diagnostic or therapeutic purposes.

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