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Freshly reconstituted guinea pig complement

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Freshly reconstituted guinea pig complement is a laboratory product that serves as a source of complement proteins. Complement proteins are a part of the immune system and play a role in various biological processes. This product is intended for use in scientific research and experiments, and its core function is to provide a source of these essential proteins.

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Lab products found in correlation

Neutravidin beads, by Thermo Fisher Scientific (4 mentions) Fish gelatin, by Boston BioProducts (3 mentions) Lsr 2 flow cytometer, by BD (3 mentions) Anti guinea pig c3 antibody conjugated to fitc, by MP Biomedicals (1 mentions) Recombinant ebov gp, by IBT Bioservices (1 mentions) Gelatin, by Boston BioProducts (1 mentions)

Close spelling variants of this product

Reconstituted guinea pig complement Guinea pig complement

4 protocols using freshly reconstituted guinea pig complement

1

Zika Virus Antibody Complement Activation Assay

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Recombinant soluble ZIKV E protein was biotinylated and coupled to red fluorescent Neutravidin beads (Life Technologies). Antibodies were diluted to 5 μg/mL in RPMI-1640, and incubated with GP-coated beads for 2 hrs at 37°C. Freshly reconstituted guinea pig complement (Cedarlane Labs) was diluted in veronal buffer with 0.1% fish gelatin (Boston Bioproducts), added to the antibody-bead complexes, and incubated for 20 min at 37°C. Beads were washed twice with PBS containing 15 mM EDTA, and stained with 1:100 dilution of an anti-guinea pig C3 antibody conjugated to Fluorescein isothiocyanate (FITC; MP Biomedicals) for 15 min at ambient temperature. Beads were washed twice more with PBS, and C3 deposition onto beads was analyzed on a BD LSRII flow cytometer and the MFI of the FITC+ of all beads was measured. Z-score values were calculated as described above.
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
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2

EBOV GP-Mediated Complement Activation

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Recombinant EBOV GP (IBT Bioservices) was biotinylated and coupled to red fluorescent Neutravidin beads (Life Technologies). Antibodies were diluted to 5μg/mL in RPMI-1640, and incubated with GP-coated beads for 2 hr at 37°C. Freshly reconstituted guinea pig complement (Cedarlane Labs) was diluted in veronal buffer with 0.1% fish gelatin (Boston Bioproducts), added to the antibody-bead complexes, and incubated for 20 min at 37°C. Beads were washed twice with phosphate buffered saline containing 15 mM EDTA, and stained with an anti-guinea pig C3 antibody conjugated to FITC (MP Biomedicals) for 15 min at ambient temperature. Beads were washed twice more with PBS, and C3 deposition onto beads was analyzed on a BD LSRII flow cytometer and the median fluorescence intensity of the FITC+ of all beads was measured.
Gilchuk P., Kuzmina N., Ilinykh P.A., Huang K., Gunn B.M., Bryan A., Davidson E., Doranz B.J., Turner H.L., Fusco M.L., Bramble M.S., Hoff N.A., Binshtein E., Kose N., Flyak A.I., Flinko R., Orlandi C., Carnahan R., Parrish E.H., Sevy A.M., Bombardi R.G., Singh P.K., Mukadi P., Muyembe-Tamfum J.J., Ohi M.D., Saphire E.O., Lewis G.K., Alter G., Ward A.B., Rimoin A.W., Bukreyev A, & Crowe JE J.r. (2018). Multifunctional Pan-ebolavirus Antibody Recognizes a Site of Broad Vulnerability on the Ebolavirus Glycoprotein. Immunity, 49(2), 363-374.e10.
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3

Zika Virus Antibody Complement Activation Assay

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Recombinant soluble ZIKV E protein was biotinylated and coupled to red fluorescent Neutravidin beads (Life Technologies). Antibodies were diluted to 5 μg/mL in RPMI-1640, and incubated with GP-coated beads for 2 hrs at 37°C. Freshly reconstituted guinea pig complement (Cedarlane Labs) was diluted in veronal buffer with 0.1% fish gelatin (Boston Bioproducts), added to the antibody-bead complexes, and incubated for 20 min at 37°C. Beads were washed twice with PBS containing 15 mM EDTA, and stained with 1:100 dilution of an anti-guinea pig C3 antibody conjugated to Fluorescein isothiocyanate (FITC; MP Biomedicals) for 15 min at ambient temperature. Beads were washed twice more with PBS, and C3 deposition onto beads was analyzed on a BD LSRII flow cytometer and the MFI of the FITC+ of all beads was measured. Z-score values were calculated as described above.
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
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4

Quantifying Anti-ZIKV NS1 Antibody Complement Activation

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Recombinant ZIKV NS1 protein (Native Antigen) was biotinylated and coupled to red fluorescent Neutravidin beads (Life Technologies) and then incubated with serial dilutions of anti-NS1 mAbs for 2 h at 37 °C to allow for binding. Freshly reconstituted guinea pig complement (Cedarlane Labs) was diluted 1 : 10 in veronal buffer containing 0.1% gelatin (Boston Bioproducts), which then was incubated with the antibody-bead complexes for 20 min at 37 °C. After washing in 15 mM EDTA in PBS, the complexes were stained with a fluorescein isothiocyanate (FITC)-conjugated anti-guinea pig C3b antibody (1 : 100; MP Biomedicals 0855385) for 15 min at room temperature. Complement deposition was detected using an IntelliCyt iQue Screener Plus or a 3 L Stratedigm S1300EXI flow cytometer, and the median fluorescent intensity of FITC was measured for all beads using FlowJo software.
Wessel A.W., Kose N., Bombardi R.G., Roy V., Chantima W., Mongkolsapaya J., Edeling M.A., Nelson C.A., Bosch I., Alter G., Screaton G.R., Fremont D.H., Crowe JE J.r, & Diamond M.S. (2020). Antibodies targeting epitopes on the cell-surface form of NS1 protect against Zika virus infection during pregnancy. Nature Communications, 11, 5278.
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