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Microtitre elisa plates

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Microtitre ELISA plates are a type of laboratory equipment used in enzyme-linked immunosorbent assay (ELISA) experiments. They consist of a flat plate with multiple individual wells that can hold small volumes of liquid samples. The plates are designed to facilitate the efficient performance of ELISA, a widely used analytical technique for the detection and quantification of various analytes, such as proteins, antibodies, or hormones, in a sample.

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Lab products found in correlation

Mouse anti his capture antibody, by Thermo Fisher Scientific (2 mentions) Alkaline phosphatase conjugated goat anti human fab secondary antibody, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

Microtitre plates ELISA plates

3 protocols using microtitre elisa plates

1

ELISA Quantification of Antibody Binding

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Antibodies b12, 2G12, PGT121, PGT128 and PGT135 were obtained from the International AIDS Vaccine Initiative (IAVI, New York, NY). Microtitre ELISA plates (Corning) were coated with mouse anti-HIS capture antibody (2 μg mL−1 in PBS; Life Technologies) overnight at 4 °C. Plates were washed five times with a solution of PBS containing 0.05% Tween 20 (v/v) and then blocked for 1 h at room temperature with 5% non-fat milk in PBS + 0.05% Tween (blocking buffer). Gp120 was then added at a concentration of 2 μg ml−1 in blocking buffer, followed by incubation for 1 h at room temperature. Plates were washed (×5) before addition of primary antibodies (b12, 17b, 2G12, PGT121, PGT128 & PGT135). A 1:5 dilution series was used starting at 20 μg ml−1 or 40 μg ml−1 in blocking buffer. Incubation was carried out at room temperature for 2 h. Plates were then washed (×5) and alkaline phosphatase-conjugated goat anti-human Fab secondary antibody (Thermo Scientific Pierce) was added as a 1:1000 dilution in blocking buffer. Plates were washed (×5) and then AP substrate (50 μL per well) was added. Once color had developed the OD at 405 nm was measured.
Pritchard L.K., Spencer D.I., Royle L., Bonomelli C., Seabright G.E., Behrens A.J., Kulp D., Menis S., Krumm S.A., Dunlop D.C., Crispin D.J., Bowden T.A., Scanlan C.N., Ward A.B., Schief W.R., Doores K.J, & Crispin M. (2015). Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. Nature communications, 6, 7479.
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2

ELISA Quantification of Antibody Binding

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Antibodies b12, 2G12, PGT121, PGT128 and PGT135 were obtained from the International AIDS Vaccine Initiative (IAVI, New York, NY). Microtitre ELISA plates (Corning) were coated with mouse anti-HIS capture antibody (2 μg mL−1 in PBS; Life Technologies) overnight at 4 °C. Plates were washed five times with a solution of PBS containing 0.05% Tween 20 (v/v) and then blocked for 1 h at room temperature with 5% non-fat milk in PBS + 0.05% Tween (blocking buffer). Gp120 was then added at a concentration of 2 μg ml−1 in blocking buffer, followed by incubation for 1 h at room temperature. Plates were washed (×5) before addition of primary antibodies (b12, 17b, 2G12, PGT121, PGT128 & PGT135). A 1:5 dilution series was used starting at 20 μg ml−1 or 40 μg ml−1 in blocking buffer. Incubation was carried out at room temperature for 2 h. Plates were then washed (×5) and alkaline phosphatase-conjugated goat anti-human Fab secondary antibody (Thermo Scientific Pierce) was added as a 1:1000 dilution in blocking buffer. Plates were washed (×5) and then AP substrate (50 μL per well) was added. Once color had developed the OD at 405 nm was measured.
Pritchard L.K., Spencer D.I., Royle L., Bonomelli C., Seabright G.E., Behrens A.J., Kulp D., Menis S., Krumm S.A., Dunlop D.C., Crispin D.J., Bowden T.A., Scanlan C.N., Ward A.B., Schief W.R., Doores K.J, & Crispin M. (2015). Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. Nature communications, 6, 7479.
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3

HIV bnAb Binding Assay via ELISA

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Microtitre ELISA plates (Corning) were coated with recombinant gp120JR-CSF (5 μg/mL in PBS) overnight at 4 °C. Plates were washed five times with a solution of PBS containing 0.05% Tween 20 (v/v) and then blocked for 1 h at room temperature with 5% non-fat milk in PBS + 0.05% Tween (blocking buffer). Plates were emptied before addition of a titration of BanLec (starting concentration, 500 μg/mL in blocking buffer using a 1:3 dilution series) and incubated for 30–60 min. HIV bnAbs were then added at a constant concentration (Ab concentration represented IC50 and were diluted in blocking buffer) and incubated for a further 1.5 h. Plates were then washed (×5) and alkaline phosphatase (AP)-conjugated goat anti-human Fab secondary antibody (Thermo Scientific Pierce) was added at a 1:1000 dilution in blocking buffer and incubated for 1 h. Plates were washed (×5) and then AP substrate (50 μL / well) was added. The OD at 405 nm was measured after 20 min.
Hopper J.T., Ambrose S., Grant O.C., Krumm S.A., Allison T.M., Degiacomi M.T., Tully M.D., Pritchard L.K., Ozorowski G., Ward A.B., Crispin M., Doores K.J., Woods R.J., Benesch J.L., Robinson C.V, & Struwe W.B. (2017). The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. Structure (London, England : 1993), 25(5), 773-782.e5.
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