High binding polystyrene 96 well plates

Manufactured by Corning

Sourced in United States

High-binding polystyrene 96-well plates are a type of laboratory equipment designed to provide a high-surface area for binding molecules, cells, or other samples. These plates feature a polystyrene surface that has been treated to enhance the adsorption of a variety of biological materials. The 96-well format allows for efficient, parallel processing of multiple samples.

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Lab products found in correlation

Opd substrate, by Thermo Fisher Scientific (3 mentions) Synergy htx microplate reader, by Agilent Technologies (2 mentions) Pbs t, by Merck Group (1 mentions) Pbs t, by Jackson ImmunoResearch (1 mentions) Ortho phenylenediamine, by Merck Group (1 mentions) Peroxidase conjugated rabbit anti human igg, by Jackson ImmunoResearch (1 mentions) O phenylenediamine dihydrochloride hrp substrate, by Merck Group (1 mentions) Prism v6, by GraphPad (1 mentions) Dulbecco s pbs, by Merck Group (1 mentions) 1 step ultra tmb elisa substrate solution, by Thermo Fisher Scientific (1 mentions) Fish gelatine, by Merck Group (1 mentions) Anti flag m2 monoclonal antibody, by Merck Group (1 mentions) Prism 8, by GraphPad (1 mentions) Duoset elisa development kit, by R&D Systems (1 mentions)

Spelling variants of this product

96-well plates (3030 citations) High-binding 96-well plates (91 citations) 96-well polystyrene plates (62 citations) High-binding plates (21 citations) Polystyrene plates (20 citations) 96-well high-binding polystyrene plates (12 citations) 96 wells (10 citations) Costar Polystyrene High Binding 96-well plates (4 citations) Polystyrene high binding plates (2 citations) Polystyrene high-binding surface 96-well plates (2 citations)

7 protocols using high binding polystyrene 96 well plates

ELISA-based Antibody Titer Assay

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Antibody titers against S. Typhi OmpC/F in sera were determined as previously described [22 (link)]. Briefly, high-binding 96-well polystyrene plates (Corning) were coated with 10 μg/ml of the protein preparation in 0.1 M carbonate-bicarbonate buffer, pH 9.5. Plates were incubated for 1 h at 37°C and then overnight at 4°C. Before use, plates were washed three times with PBS containing 0.05% Tween-20 (PBS-T) (Sigma–Aldrich). Non-specific binding was blocked with 5% non-fat dry milk diluted in PBS (PBS-M) for 1 h at 37°C. After washing, sera were diluted 1:40 and stool extracts 1:2, both in PBS-M and twofold serial dilutions were added to the wells. Plates were incubated for 1 h at 37°C, followed by four washes with PBS-T. After 1 h of incubation at 37°C with peroxidase-conjugated rabbit anti-human IgG (1:10,000) or IgM (1:5000) antibody (in PBS-M, Jackson Immuno Research), four washes with PBS-T, ortho-phenylenediamine (0.5 mg/ml; Sigma) in 0.1 M citrate buffer, pH 5.6, containing 0.08% H₂O₂ was used to develop the reaction. Optical density was read at 492 nm using an automated ELISA plate reader (Tecan). Antibody titers are given as -log2 dilution × 40. Antibody titers were defined as the highest dilution of the sample at which the OD was higher than the mean ± 3 SD of the negative sample values.

Carreño J.M., Perez-Shibayama C., Gil-Cruz C., Lopez-Macias C., Vernazza P., Ludewig B, & Albrich W.C. (2017). Evolution of Salmonella Typhi outer membrane protein-specific T and B cell responses in humans following oral Ty21a vaccination: A randomized clinical trial. PLoS ONE, 12(6), e0178669.

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Quantifying Biotinylated scFv Immobilization

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The effectiveness of immobilization of the biotinylated scFvs was quantified using an enzyme-linked immunosorbent assay (ELISA). High-binding 96-well polystyrene plates (Corning) were coated with streptavidin (NEB), as previously described [13 (link)]. Buffer-exchanged cell lysates were serially diluted in PBS with 0.1% Tween 20 (PBST) with 0.5% milk and loaded onto streptavidin-coated plates (50 μL/well), which had been washed with PBST prior to loading of the samples. After 45 min of incubation, unbound scFvs were washed off with PBST, and the immobilized scFvs were incubated for 1 h with an anti-FLAG antibody (HRP-conjugated, Sigma). Unbound antibody was washed off, and 200 μL of o-phenylenediamine dihydrochloride HRP substrate (Sigma) was added to each well to react with the HRP on the antibody. After the reaction proceeded for 30 minutes, 50 μL of 3 M H₂SO₄ was added to quench the reaction. The level of immobilization was quantified using absorbance at 492 nm, measured on a plate reader (Epoch, Bio-Tek). Three separate samples were evaluated on different days, with at least five replicates for each construct. For statistical analysis, two-way ANOVA tests (p<0.05) with the Bonferroni correction for multiple comparison against no linker constructs were done at 1 μg/μL, 0.063 μg/μL, and 0.008 μg/μL total protein concentrations.

Ikonomova S.P., Le M.T., Kalla N, & Karlsson A.J. (2018). Effect of linkers on immobilization of scFvs with biotin-streptavidin interaction. Biotechnology and applied biochemistry, 65(4), 580-585.

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ZIKV Antibody Quantification by ELISA

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Plasma anti-ZIKV antibody concentrations were measured by end point dilution ELISA. For this assay, high-binding polystyrene 96-well plates (Corning) were coated with PBS containing a 1:1000 dilution of 4x10⁸ ffu/ml stock of purified ZIKV particle preparations The plates were incubated overnight at 4°C and then blocked with PBS containing 2% milk and 0.05% Tween (ELISA-Block) for 1 hr at room temperature. Plates were washed with 0.05% Tween-PBS (ELISA-Wash) and incubated with two-fold dilutions of RM plasma in ELISA-Block starting at a dilution of 1:50. The plate was incubated at room temperature for 2 hrs. Plates were washed several times with ELISA-Wash and then incubated with secondary anti-monkey IgM or IgG (Rockland, Inc.) conjugated with horseradish peroxidase for 30 mins. Plates were washed with ELISA-Wash and bound secondary antibody was detected using the OPD substrate (Life Technologies) followed by HCl stop the assay. The plates were read within 10 minutes using a Synergy HTX Microplate Reader (BioTek) at 490nm. Endpoint titers of ZIKV binding antibodies were determined using a Log/Log transformation method and the results were analyzed and graphed using GraphPad Prism v6 software.

Hirsch A.J., Smith J.L., Haese N.N., Broeckel R.M., Parkins C.J., Kreklywich C., DeFilippis V.R., Denton M., Smith P.P., Messer W.B., Colgin L.M., Ducore R.M., Grigsby P.L., Hennebold J.D., Swanson T., Legasse A.W., Axthelm M.K., MacAllister R., Wiley C.A., Nelson J.A, & Streblow D.N. (2017). Zika Virus infection of rhesus macaques leads to viral persistence in multiple tissues. PLoS Pathogens, 13(3), e1006219.

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ZIKV Antibody Quantification by ELISA

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Anti-ZIKV antibody levels were measured in maternal and fetal blood plasma by end point dilution ELISA as previously described [28 (link)]. Briefly, high-binding polystyrene 96-well plates (Corning) were coated with purified ZIKV particle preparations overnight at 4°C. The plates were blocked with PBS containing 2% milk and 0.05% Tween (ELISA-Block) for 1 hr at room temperature. Plates were washed with ELISA-Wash and incubated with two-fold dilutions of plasma samples in ELISA-Block for 2 hrs. Plates were washed with ELISA-Wash and then incubated for 30 mins with secondary anti-monkey IgM or IgG (Rockland, Inc.) conjugated with horseradish peroxidase. Bound secondary antibody was detected using the OPD substrate (Life Technologies) followed by addition of HCl to stop the reaction. The plates were read using a Synergy HTX Microplate Reader (BioTek) at 490nm. Endpoint antibody binding titers were calculated by Log/Log transformation and analyzed using GraphPad Prism v6 software. The limit of detection was a dilution = 1:50.

Steinbach R.J., Haese N.N., Smith J.L., Colgin L.M., MacAllister R.P., Greene J.M., Parkins C.J., Kempton J.B., Porsov E., Wang X., Renner L.M., McGill T.J., Dozier B.L., Kreklywich C.N., Andoh T.F., Grafe M.R., Pecoraro H.L., Hodge T., Friedman R.M., Houser L.A., Morgan T.K., Stenzel P., Lindner J.R., Schelonka R.L., Sacha J.B., Roberts V.H., Neuringer M., Brigande J.V., Kroenke C.D., Frias A.E., Lewis A.D., Kelleher M.A., Hirsch A.J, & Streblow D.N. (2020). A neonatal nonhuman primate model of gestational Zika virus infection with evidence of microencephaly, seizures and cardiomyopathy. PLoS ONE, 15(1), e0227676.

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SARS-CoV-2 RBD Binding Assay

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High binding polystyrene 96-well plates (Corning Inc., Corning, NY, USA) were coated with 2 µg/ml streptavidin (Thermo Fisher Scientific, Waltham, MA, USA) diluted in Dulbecco’s PBS (HyClone, South Logan, UT, USA) over night at +4°C. On the following day the plate was washed and subsequently incubated for 30 min with 30µl 30 nM biotinylated SARS-CoV-2 RBD (SinoBiological; product number:40592-V27H-B) diluted in Dulbecco’s PBS containing 0.05% Tween 20 and 0.5% fish gelatine (Sigma Aldrich, St. Louis, MO, USA) (assay buffer). After washing the immobilized antigen was preincubated for 40 minutes at room temperature with 30 µl assay buffer or assay buffer containing 4.8 pmol IgG. Subsequently, 10 µl of assay buffer or assay buffer containing 4.8 pmol scFv was added to each well. After 1 hour incubation at room temperature the wells were washed and bound scFv was detected by incubation for 40 minutes at room temperature with peroxidase labelled monoclonal anti-FLAG^® M2 antibody (Sigma Aldrich (30 µl diluted 1/4000 in assay buffer) and development using 1-Step™ Ultra TMB-ELISA Substrate Solution (Thermo Fisher Scientific).

Bahnan W., Wrighton S., Sundwall M., Bläckberg A., Larsson O., Höglund U., Khakzad H., Godzwon M., Walle M., Elder E., Strand A.S., Happonen L., André O., Ahnlide J.K., Hellmark T., Wendel-Hansen V., Wallin R.P., Malmstöm J., Malmström L., Ohlin M., Rasmussen M, & Nordenfelt P. (2022). Spike-Dependent Opsonization Indicates Both Dose-Dependent Inhibition of Phagocytosis and That Non-Neutralizing Antibodies Can Confer Protection to SARS-CoV-2. Frontiers in Immunology, 12, 808932.

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ZIKV Antibody Detection in Maternal-Fetal Samples

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Anti-ZIKV antibodies were measured by ELISA in the maternal and fetal plasma as well as in the cord blood (venous and arterial) and amniotic fluid. High-binding polystyrene 96-well plates (Corning) were coated overnight with 100 µl of PBS containing a 1:100 dilution of purified ZIKV particle preparations. The plates were blocked with PBS containing 2% milk and 0.05% Tween (ELISA-Block) for 1 h at room temperature, washed with 0.05% Tween-PBS (ELISA-Wash), and incubated with two-fold dilutions of rhesus plasma in ELISA-Block starting at a dilution of 1:50. The plate was incubated at room temperature for 2 h. Plates were washed several times with ELISA-Wash and then incubated with secondary anti-monkey IgM/A/G (Rockland, Inc.) conjugated with horseradish peroxidase for 30 mins. Plates were washed with ELISA-Wash and bound secondary antibody was detected using the OPD substrate (Life Technologies) with an HCl stop. The plates were read within 10 min using a Synergy HTX Microplate Reader (BioTeck) at 490 nm. Dilution titers of ZIKV-binding antibodies were determined using a Log-to-Log transformation method and the results were graphed using GraphPad Prism v6 software.

Hirsch A.J., Roberts V.H., Grigsby P.L., Haese N., Schabel M.C., Wang X., Lo J.O., Liu Z., Kroenke C.D., Smith J.L., Kelleher M., Broeckel R., Kreklywich C.N., Parkins C.J., Denton M., Smith P., DeFilippis V., Messer W., Nelson J.A., Hennebold J.D., Grafe M., Colgin L., Lewis A., Ducore R., Swanson T., Legasse A.W., Axthelm M.K., MacAllister R., Moses A.V., Morgan T.K., Frias A.E, & Streblow D.N. (2018). Zika virus infection in pregnant rhesus macaques causes placental dysfunction and immunopathology. Nature Communications, 9, 263.

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Quantifying TNF-α Release by ELISA

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The amount of released pro-inflammatory cytokine TNF-α was determined using a DuoSet ELISA Development Kit (R&D Systems, Zug, Switzerland) according to the supplier’s protocols. For one sample (LPS 24 h), the samples were diluted 1:5 (v/v) in reagent diluent to remain within the detection limit of the instrument. The measurements were performed in high-binding polystyrene 96-well plates (Corning). Standards and samples were run in triplicates. The concentrations of the TNF-α released in the cell culture medium were calculated based on the standard curves and fitted with a four-parameter logistic (4PL) approach using GraphPad Prism 8 software (GraphPad Software Inc., San Diego, CA, USA).

Susnik E., Taladriz-Blanco P., Drasler B., Balog S., Petri-Fink A, & Rothen-Rutishauser B. (2020). Increased Uptake of Silica Nanoparticles in Inflamed Macrophages but Not upon Co-Exposure to Micron-Sized Particles. Cells, 9(9), 2099.

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