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Sβg diluent

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SβG Diluent is a laboratory reagent used to dilute samples during the analysis of serum beta-glucuronidase (SβG) levels. It provides a consistent and controlled dilution environment for accurate quantification of SβG in various sample types.

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Lab products found in correlation

Hd x analyzer, by Quanterix (4 mentions) Streptavidin β galactosidase concentrate, by Quanterix (3 mentions) Homebrew detector sample diluent, by Quanterix (2 mentions) Bead diluent, by Quanterix (1 mentions)

4 protocols using sβg diluent

1

Simoa Assay Protocol for Multiplexed Protein Quantification

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All Simoa assays were performed on an HD-X Analyzer (Quanterix), with automated sample processing, image analysis, and calculations of average enzymes per bead (AEB). Assay conditions for each analyte are listed in Table S12. The same antibody-coated beads used in the single-plex MOSAIC assays were used for all Simoa assays, with the same detector antibody concentrations as in the MOSAIC assays. For each single-plex Simoa assay, 100 000 antibody-coated beads and 400 000 helper (nonconjugated) beads were used; for each four-plex Simoa assay, 125 000 antibody-coated beads per analyte were used. Streptavidin-β-galactosidase (SβG) Concentrate (Quanterix) was diluted in SβG Diluent (Quanterix) to the desired concentration for each assay. All assay reagents and consumables were loaded to the HD-X Analyzer according to the manufacturer’s instructions.
Wu C., Dougan T.J, & Walt D.R. (2022). High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity. ACS nano, 16(1), 1025-1035.
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2

Conventional Simoa Assay Workflow

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Conventional Simoa assays were performed on an HD-X Analyzer (Quanterix), using the same antibody-coated capture beads (500000 beads per assay) and biotinylated detector antibodies at the same concentrations as in the corresponding dSimoa assays and 100 μL sample volumes. Streptavidin-β-galactosidase (SβG) Concentrate (Quanterix) was diluted in SβG Diluent (Quanterix) to the desired concentration. The same incubation time of 1 h was used for the antibody capture step in which the beads, sample, and detector antibody are incubated for immunocomplex sandwich formation. For each target, two assay conditions were performed: one assay with the same SβG concentrations and incubation times as in the corresponding dSimoa assay, and one assay with a standard SβG concentration and incubation time used on the HD-X (150 pM SβG for 5 min). Beads, detector antibody, and SβG were placed in plastic bottles (Quanterix), and samples were added to a 96-well plate, all of which were loaded into the HD-X Analyzer. The enzyme substrate (resorufin β-D-galactopyranoside), Wash Buffer 1, Wash Buffer 2, and Simoa Sealing Oil were loaded into the HD-X Analyzer according to the manufacturer’s instructions. All assay steps, image analyses, and calculations of average enzyme per bead (AEB) were automated, as previously described in detail.13
Wu C., Garden P.M, & Walt D.R. (2020). Ultrasensitive Detection of Attomolar Protein Concentrations by Dropcast Single Molecule Assays. Journal of the American Chemical Society, 142(28), 12314-12323.
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3

SARS-CoV-2 Serological Simoa Assays

Cited 1 time
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SARS-CoV-2 serological Simoa assays for IgG against four viral antigen spike 1 subunit (S1), spike (stabilized ectodomain of spike with mutated furin cleavage site), Nucleocapsid, and RBD were prepared and preformed as previously described.32 (link) Briefly, plasma samples were diluted 4000-fold in Homebrew Detector/Sample Diluent (Quanterix Corp.). Four antigen-conjugated capture beads were mixed and diluted in Bead Diluent, with a total of 500,000 beads per reaction (125,000 of each bead type). Biotinylated anti-human IgG antibodies (Bethyl Laboratories A80-148B) were diluted in Homebrew Detector/Sample Diluent to final concentrations of 7.73ng/mL. Streptavidin-β-galactosidase (SβG) was diluted to 30 pM in SβG Diluent (Quanterix). The serology assay was performed on an HD-X Analyzer (Quanterix) in an automated three-step assay. Average Enzyme per Bead (AEB) values were calculated by the HD-X Analyzer software. All samples were measured in duplicates.
Mysore V., Cullere X., Settles M.L., Ji X., Kattan M.W., Desjardins M., Durbin-Johnson B., Gilboa T., Baden L.R., Walt D.R., Lichtman A.H., Jehi L, & Mayadas T.N. (2021). Protective heterologous T cell immunity in COVID-19 induced by the trivalent MMR and Tdap vaccine antigens. Med (New York, N.y.), 2(9), 1050-1071.e7.
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4

Simoa Serology Assay for SARS-CoV-2 Antibodies

Cited 1 time
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Simoa assays for IgG, IgA, and IgM against four SARS-CoV-2 targets (spike, S1, nucleocapsid, and RBD) were performed as previously described (Norman et al., 2020 (link)). Reference materials were diluted 1:250-, 1:1,000-, 1:4,000-, and 1:16,000-fold in Homebrew Detector/Sample Diluent (Quanterix Corporation, Product code: 101359, Billerica, Massachusetts, USA). Four antigen-conjugated capture beads were mixed and diluted in Bead Diluent (Quanterix Corporation, Product code: 101362, Billerica, Massachusetts, USA), with a total of 500,000 beads per reaction (125,000 of each bead type). Biotinylated antibodies were diluted in Homebrew Detector/Sample Diluent to final concentrations of IgG (Bethyl Laboratories A80-148B; Montgomery, Texas, USA): 7.73 ng/ml, IgA (Abcam ab214003, Waltham, Massachusetts, USA): 150 ng/ml, and IgM (Thermo Fisher Scientific, MII0401, Pittsburgh, Pennsylvania, USA): 216 ng/ml: Streptavidin-β-galactosidase (SβG) concentrate (Quanterix Corporation, Product code: 1013397, Billerica, Massachusetts, USA) was diluted to 30 pM in SβG Diluent (Quanterix Corporation, Product code: 100376, Billerica, Massachusetts, USA). The serology assay was performed on the HD-X Analyzer (Quanterix) in an automated three-step assay. Average enzymes per bead (AEB) values were calculated by the HD-X Analyzer software (Norman et al., 2020 (link)).
Windsor W.J., Roell Y., Tucker H., Cheng C.A., Suliman S., Peek L.J., Pestano G.A., Lee W.T., Zeichhardt H., Lamb M.M., Kammel M., Wang H., Kedl R., Rester C., Morrison T.E., Davenport B.J., Carson K., Yates J., Howard K., Kulas K., Walt D.R., Dafni A., Taylor D, & Chu M. (2022). Harmonization of Multiple SARS-CoV-2 Reference Materials Using the WHO IS (NIBSC 20/136): Results and Implications. Frontiers in Microbiology, 13, 893801.
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