Calcium and magnesium

Manufactured by Boston BioProducts

Sourced in Canada

Calcium and magnesium are essential mineral nutrients commonly used in laboratory settings. These elements play crucial roles in various biological processes. Calcium is involved in cellular signaling, bone structure, and muscle function, while magnesium is required for energy production, protein synthesis, and nerve function. This product provides a reliable source of these important minerals for laboratory applications.

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Lab products found in correlation

Red neutravidin conjugated microspheres, by Thermo Fisher Scientific (3 mentions) Lyophilized guinea pig complement, by Cedarlane (3 mentions) Sulfo nhs, by Thermo Fisher Scientific (2 mentions) Carboxylate modified microspheres, by Thermo Fisher Scientific (2 mentions) Guinea pig complement ce, by MP Biomedicals (2 mentions) Reconstituted guinea pig complement, by Cedarlane (1 mentions) Fitc conjugated anti guinea pig c3 antibody, by MP Biomedicals (1 mentions) Ez link sulfo nhs lc lc, by Thermo Fisher Scientific (1 mentions) Gelatin veronal buffer, by Boston BioProducts (1 mentions)

4 protocols using calcium and magnesium

ADCD Assay for Complement Activation

Cited 1 time

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ADCD was conducted as previously described^{58 (link)}. Briefly, NVX-CoV2373 Spike protein was biotinylated using EDC (Thermo Fisher) and Sulfo-NHS (Thermo Fisher), and then coupled to red Neutravidin-conjugated microspheres (Thermo Fisher) or directly coupled to Carboxylate-Modified microspheres (Thermo Fisher). Immune complexes were formed by incubating the bead + protein conjugates with diluted serum for 2 hours at 37°C, and then washed to remove unbound antibody. The immune complexes were then incubated with lyophilized guinea pig complement (Cedarlane) and diluted in gelatin veronal buffer with calcium and magnesium (Boston Bioproducts) for 30 minutes. C3 bound to immune complexes was detected by fluorescein-conjugated goat IgG fraction to guinea pig Complement Ce (MP Biomedicals). Flow cytometry was performed to identify the percentage of beads with bound C3. Flow cytometry was performed with an IQue (Intellicyt) and analysis was performed on IntelliCyt ForeCyt (v8.1).

Alter G., Gorman M., Patel N., Guebre-Xabier M., Zhu A., Atyeo C., Pullen K., Loos C., Goez-Gazi Y., Carrion R J.r., Tian J.H., Yuan D., Bowman K., Zhou B., Maciejewski S., McGrath M., Logue J., Frieman M., Montefiori D., Schendel S., Saphire E.O., Lauffenburger D., Greene A., Portnoff A., Massare M., Ellingsworth L., Glenn G., Smith G., Mann C., Amanat F, & Krammer F. (2021). Collaboration between the Fab and Fc contribute to maximal protection against SARS-CoV-2 following NVX-CoV2373 subunit vaccine with Matrix-M™ vaccination. Research Square.

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Complement-dependent Antibody Interaction Assay

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This assay is designed to replicate in vivo interaction of complement/antibody complexes with free antigens and/or virus. Recombinant EBOV GPΔTM was biotinylated and coupled to red Neutravidin beads (ThermoScientific) as described above for ADCP. Heat-inactivated sera was diluted 1:10 in culture medium and incubated with GP-coated beads for 2h at 37°C. Unbound antibodies were removed by centrifugation prior to the addition of reconstituted guinea pig complement (Cedarlane Labs) diluted in veronal buffer supplemented with calcium and magnesium (Boston Bioproducts) for 20 min at 37°C. Beads were washed with PBS containing 15mM EDTA, and stained with an FITC-conjugated anti-guinea pig C3 antibody (MP Biomedicals, Santa Ana, CA). C3 deposition onto beads was analyzed by flow cytometry. The gMFI of FITC of all beads was measured.

Gunn B.M., McNamara R.P., Wood L., Taylor S., Devadhasan A., Guo W., Das J., Nilsson A., Shurtleff A., Dubey S., Eichberg M., Suscovich T.J., Saphire E.O., Lauffenburger D., Coller B.A., Simon J.K, & Alter G. (2023). Antibodies against the Ebola virus soluble glycoprotein are associated with long-term vaccine-mediated protection of non-human primates. Cell reports, 42(4), 112402.

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Antibody-Dependent Complement Deposition Assay

Cited 1 time

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ADCD was conducted as previously described^{58 (link)}. Briefly, NVX-CoV2373 Spike protein was biotinylated using EDC (Thermo Fisher) and Sulfo-NHS (Thermo Fisher), and then coupled to red Neutravidin-conjugated microspheres (Thermo Fisher) or directly coupled to Carboxylate-Modified microspheres (Thermo Fisher). Immune complexes were formed by incubating the bead+protein conjugates with diluted serum for 2 hours at 37°C, and then washed to remove unbound antibody. The immune complexes were then incubated with lyophilized guinea pig complement (Cedarlane) and diluted in gelatin veronal buffer with calcium and magnesium (Boston Bioproducts) for 30 minutes. C3 bound to immune complexes was detected by fluorescein-conjugated goat IgG fraction to guinea pig Complement Ce (MP Biomedicals). Flow cytometry was performed to identify the percentage of beads with bound C3. Flow cytometry was performed with an IQue (Intellicyt) and analysis was performed on IntelliCyt ForeCyt (v8.1).

Gorman M.J., Patel N., Guebre-Xabier M., Zhu A., Atyeo C., Pullen K.M., Loos C., Goez-Gazi Y., Carrion R J.r., Tian J.H., Yaun D., Bowman K., Zhou B., Maciejewski S., McGrath M.E., Logue J., Frieman M.B., Montefiori D., Mann C., Schendel S., Amanat F., Krammer F., Saphire E.O., Lauffenburger D., Greene A.M., Portnoff A.D., Massare M.J., Ellingsworth L., Glenn G., Smith G, & Alter G. (2021). Collaboration between the Fab and Fc contribute to maximal protection against SARS-CoV-2 in nonhuman primates following NVX-CoV2373 subunit vaccine with Matrix-M™ vaccination. bioRxiv.

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Complement Deposition Assay for Antibodies

Cited 1 time

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ADCD was conducted as previously described [82 (link)]. Briefly, spike or RBD protein was biotinylated using EDC (Thermo Fisher Scientific) and EZ-link Sulfo-NHS-LC-LC (Thermo Fisher Scientific) and then coupled to red Neutravidin-conjugated microspheres (Thermo Fisher Scientific). Immune complexes were formed by incubating the bead+protein conjugates with diluted serum (ADCD 1:10 dilution) for 2 hours at 37°C and then washed to remove unbound antibody. The immune complexes were then incubated with lyophilized guinea pig complement (Cedarlane - Burlington, Ontario, Canada) and diluted in gelatin veronal buffer with calcium and magnesium (Boston BioProducts - Milford, Massachusetts, USA) for 30 minutes. C3 bound to immune complexes was detected by fluorescein-conjugated goat IgG fraction to guinea pig Complement C3 (MP Biomedicals - Irvine, California, USA). Flow cytometry was performed to identify the percentage of beads with bound C3 and a complement deposition score was calculated (% beads positive × median fluorescent intensity of positive beads). Flow cytometry was performed with an LSRII (BD) and analysis was performed using FlowJo V10.7.1.

Zhu D.Y., Gorman M.J., Yuan D., Yu J., Mercado N.B., McMahan K., Borducchi E.N., Lifton M., Liu J., Nampanya F., Patel S., Peter L., Tostanoski L.H., Pessaint L., Van Ry A., Finneyfrock B., Velasco J., Teow E., Brown R., Cook A., Andersen H., Lewis M.G., Lauffenburger D.A., Barouch D.H, & Alter G. (2022). Defining the determinants of protection against SARS-CoV-2 infection and viral control in a dose-down Ad26.CoV2.S vaccine study in nonhuman primates. PLoS Biology, 20(5), e3001609.

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