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High binding elisa plates

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High binding ELISA plates are a type of laboratory equipment used for enzyme-linked immunosorbent assay (ELISA) applications. These plates feature a high-affinity surface that enhances the binding of proteins or other biomolecules, allowing for efficient capture and detection in ELISA experiments.

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Lab products found in correlation

Goat anti mouse igg2b, by Southern Biotech (3 mentions) Tmb substrate, by Thermo Fisher Scientific (2 mentions) Peroxidase conjugated goat anti mouse igg h l, by Jackson ImmunoResearch (2 mentions) Neutralite avidin peroxidase n hrp, by Southern Biotech (2 mentions) Triton x, by Merck Group (2 mentions) 1 step ultra tmb elisa substrate solution, by Thermo Fisher Scientific (2 mentions) Nb200 540b, by Novus Biologicals (2 mentions) Streptavidin hrp, by Mabtech (2 mentions) Triton x 100 detergent, by Merck Group (1 mentions) Neutralite avidin peroxidase, by Southern Biotech (1 mentions) Infinite m200, by Tecan (1 mentions) Serum gel, by Sarstedt (1 mentions)

Close spelling variants of this product

MaxiSorp high-binding 96-well ELISA plates MaxiSorp high binding ELISA plates Nunc MaxiSorp high protein-binding capacity 96-well ELISA plates 96-well high-binding ELISA plates High-binding plates ELISA plates ELISAs

9 protocols using high binding elisa plates

1

Mouse IgG Antibody ELISA Assay

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High-binding ELISA plates (eBioscience) were coated with 20 μg/mL of antigen (KFE8-Ag85B) in PBS overnight at 4 °C and blocked with 200 μL of 1% BSA in PBST (0.5% Tween-20 in PBS) for 1 h. Serum dilutions were applied (1:10−2 to 1:10−9, 100 μL/well) for 1 h at room temperature followed by addition of peroxidase-conjugated goat anti-mouse IgG (H + L) (Jackson Immuno Research) (1:5000 in 1% BSA-PBST, 100 μL/well). Plates were developed using TMB substrate (100 μL/well, eBioscience), the reaction stopped using 50 μl of 1 M phosphoric acid, and absorbance measured at 450 nm. Absorbance values of PBS (no antigen) coated wells were subtracted to account for background.
Chesson C.B., Huante M., Nusbaum R.J., Walker A.G., Clover T.M., Chinnaswamy J., Endsley J.J, & Rudra J.S. (2018). Nanoscale Peptide Self-assemblies Boost BCG-primed Cellular Immunity Against Mycobacterium tuberculosis. Scientific Reports, 8, 12519.
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2

CocKFE8 Nanofiber ELISA Antibody Assay

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High-binding ELISA plates (eBioscience, San Diego, CA) were coated with 20 μg/mL of CocKFE8 nanofibers in PBS overnight at 4 °C and blocked with 200 μL of 1% BSA in PBST (0.5% Tween-20 in PBS) for 1 h. Serum was diluted in PBST (1:100) and applied (100 μL/well) for 1 h at room temperature followed by peroxidase-conjugated goat anti-mouse IgG (H+L) (Jackson Immuno Research, West Grove, PA) (1:5000 in 1% BSA-PBST, 100 μL/well). Plates were developed using TMB substrate (100 μL/well, eBioscience, San Diego, CA), the reaction stopped using 50 μL of 1 M phosphoric acid, and absorbance measured at 450 nm. Absorbance values of PBS (no antigen)-coated wells were subtracted to account for background. Vaccinated mice with titers lower than three times the standard deviation of the mean titer for the control group (PBS-treated; titers lower than 0.05 absorbance units) were defined as “CocKFE8 nonresponders” and all other vaccinated mice were defined as “CocKFE8 responders” and compared to control mice in the behavioral studies.
Rudra J.S., Ding Y., Neelakantan H., Ding C., Appavu R., Stutz S., Snook J.D., Chen H., Cunningham K.A, & Zhou J. (2016). Suppression of Cocaine-Evoked Hyperactivity by Self-Adjuvanting and Multivalent Peptide Nanofiber Vaccines. ACS chemical neuroscience, 7(5), 546-552.
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3

Quantitative SIV p27 ELISA Assay

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High binding ELISA plates (ThermoFisher, Waltham, MA) were coated at 1μg/mL with goat anti-mouse IgG2b (Southern Biotech, Birmingham, AL) overnight at 4°C. The next day, plates were washed 6x with 0.05% PBS-tween (PBST) and blocked with 1% BSA in PBST for 30 minutes at RT. Mouse anti-p27 2F12 antibody (AIDS reagent Program), at 1μg/mL, was added to plates and incubated for 1 hr at 37°C. After washing plates 6x, p27 standard (Immune Technologies) and culture medium treated with 0.5% Triton X-100 detergent (Sigma) to lyse virus particles were added in 3-fold dilutions and allowed to incubate for 1.5 hr at 37°C. Plates were washed and biotinylated SIV IG diluted 1:1000 (prepared in the laboratory) and added to each well. After 1 hr at 37°C, plates were washed, and 1:4000 neutralite-avidin peroxidase (Southern Biotech) was added. After 30 min at RT in the dark, bound IgG was detected using tetramethylbenzidine substrate (KPL, Gaithersburg, MD). The reaction was stopped by adding 100 μl of 2N H2SO4.
Styles T.M., Gangadhara S., Reddy P.B., Sahoo A., Shiferaw A., Welbourn S., Kozlowski P.A., Derdeyn C.A., Velu V, & Amara R.R. (2022). V2 hotspot optimized MVA vaccine expressing stabilized HIV-1 Clade C envelope Gp140 delays acquisition of heterologous Clade C Tier 2 challenges in Mamu-A*01 negative Rhesus Macaques. Frontiers in Immunology, 13, 914969.
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4

ELISA Quantification of Ranibizumab

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The concentration of ranibizumab was analyzed by the ELISA method, slightly modified from previous reports34 (link). High binding ELISA plates (Thermo Scientific, USA) were coated with 10 µg/ml of VEGF165 in 50 mM carbonate buffer pH 9.6, and incubated for 24 hours at 4 °C. The plates were washed three times with 0.05% Tween-20 in PBS, and the wells were blocked with 2% BSA (0.05% Tween-20 in PBS) overnight at 4 °C. After this, the plates were washed three times, and were ready for use. Appropriately diluted ranibizumab samples were aliquoted into the wells and incubated at RT for 2 hours with gentle shaking. The plate was washed five times with 0.05% Tween-20 in PBS pH 7.4 to remove unbound protein before loading the Peroxidase conjugated goat secondary antibody anti-human IgG (diluted 1:20,000 in PBS) which was incubated for 45 min at RT. The plates were again washed five times with 0.05% Tween-20 in PBS pH 7.4 and finally, ELISA chemiluminescent substrate was added. The reading was taken by measuring the luminescence of samples using a microplate reader (TECAN infinite M200, Switzerland). A standard calibration curve was included in every ELISA plate to account for any plate-to-plate variations. The detection limit of this assay was approximately 1.5 ng/ml. Results were represented as an average of triplicate measurements.
Joseph R.R., Tan D.W., Ramon M.R., Natarajan J.V., Agrawal R., Wong T.T, & Venkatraman S.S. (2017). Characterization of liposomal carriers for the trans-scleral transport of Ranibizumab. Scientific Reports, 7, 16803.
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5

Quantification of Anti-SIV Antibodies

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High binding ELISA plates (Thermo Scientific, catalog no 44-2404-21) were coated at 0.5μg/mL with goat anti-mouse IgG2b (Southern Biotech, catalog no 1090–05) overnight at 4°C. Next day, plates were washed 6x with PBST (PBS + 0.05%Tween-20) and blocked with 1% BSA in PBST for 30 minutes at RT. Anti-p27 2F12 (NIH AIDS Reagent Program) antibody at 0.5 μg/mL was added to ELISA plates and incubated for 1 hr at 37°C. Plates were again washed 6x with PBST. Supernatant from ADCVI assays which were placed in the 4°C the night before, were treated with TritonX (Sigma) to a make a 0.5% Triton X solution to inactivate virus particles and release Gag. Gag supernatant was diluted 3-fold and added to ELISA plates and allowed to incubate for 1½ hr at 37°C. Plates were washed and biotinylated anti-SIV IgG diluted 1:1000 and added to each well. Plates were again incubated for 1 hr at 37°C. After 6x wash, neutralite-avidin peroxidase (N-HRP) (Southern Biotech) was diluted 1:4000 and added to each plate for 30 minutes at RT in the dark. After incubation, bound IgG was detected using tetramethylbenzidine substrate (KPL, Gaithersburg, MD). The reaction was stopped by adding 100 μl 2N H2SO4. The readings were recorded at 450nm.
Sahoo A., Hodge E.A., LaBranche C.C., Styles T.M., Shen X., Cheedarla N., Shiferaw A., Ozorowski G., Lee W.H., Ward A.B., Tomaras G.D., Montefiori D.C., Irvine D.J., Lee K.K, & Amara R.R. (2022). Structure-guided changes at the V2 apex of HIV-1 clade C trimer enhance elicitation of autologous neutralizing and broad V1V2-scaffold antibodies. Cell reports, 38(9), 110436.
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6

Quantification of Anti-SIV Antibodies

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High binding ELISA plates (Thermo Scientific, catalog no 44-2404-21) were coated at 0.5μg/mL with goat anti-mouse IgG2b (Southern Biotech, catalog no 1090–05) overnight at 4°C. Next day, plates were washed 6x with PBST (PBS + 0.05%Tween-20) and blocked with 1% BSA in PBST for 30 minutes at RT. Anti-p27 2F12 (NIH AIDS Reagent Program) antibody at 0.5 μg/mL was added to ELISA plates and incubated for 1 hr at 37°C. Plates were again washed 6x with PBST. Supernatant from ADCVI assays which were placed in the 4°C the night before, were treated with TritonX (Sigma) to a make a 0.5% Triton X solution to inactivate virus particles and release Gag. Gag supernatant was diluted 3-fold and added to ELISA plates and allowed to incubate for 1½ hr at 37°C. Plates were washed and biotinylated anti-SIV IgG diluted 1:1000 and added to each well. Plates were again incubated for 1 hr at 37°C. After 6x wash, neutralite-avidin peroxidase (N-HRP) (Southern Biotech) was diluted 1:4000 and added to each plate for 30 minutes at RT in the dark. After incubation, bound IgG was detected using tetramethylbenzidine substrate (KPL, Gaithersburg, MD). The reaction was stopped by adding 100 μl 2N H2SO4. The readings were recorded at 450nm.
Sahoo A., Hodge E.A., LaBranche C.C., Styles T.M., Shen X., Cheedarla N., Shiferaw A., Ozorowski G., Lee W.H., Ward A.B., Tomaras G.D., Montefiori D.C., Irvine D.J., Lee K.K, & Amara R.R. (2022). Structure-guided changes at the V2 apex of HIV-1 clade C trimer enhance elicitation of autologous neutralizing and broad V1V2-scaffold antibodies. Cell reports, 38(9), 110436.
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7

Quantifying Complement Binding via ELISA

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High binding ELISA plates (07-200-37, Fisher Scientific) were coated with 1 μg/mL of labeled eOD-60mer40 and blocked with 2% BSA in PBS. Serum was collected from naïve mice using collection tube with Serum Gel (41.1500.005, Sarstedt), diluted in PBS (3% v/v), and incubated in ELISA plates for 1 hour at 37°C. Complement binding was detected by biotinylated anti-C3 antibody (NB200-540B, Novus Biologicals) followed by streptavidin-HRP (3310-9-1000, Mabtech AB). The plates were developed with 1-Step Ultra TMB-ELISA Substrate Solution (34028, Thermo Fisher Scientific) and the reaction was stopped by adding with 2 N sulfuric acid (BDH7500-1).
Tavernier J., Tuypens T., Verhee A., Plaetinck G., Devos R., Van der Heyden J., Guisez Y, & Oefner C. (1995). Identification of receptor-binding domains on human interleukin 5 and design of an interleukin 5-derived receptor antagonist.. Proceedings of the National Academy of Sciences of the United States of America, 92(11), 5194-5198.
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8

Enzyme-Linked Immunosorbent Assay for Serum Antibody Responses

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Blood collected from immunized mice through retro-orbital bleeding was processed in collection tube with separator gel (41.1500.005, Sarstedt) to obtain serum samples. To analyze on-target antibody response, high binding ELISA plates (07-200-37, Fisher Scientific) were coated with 1 μg/mL trimer and blocked with 2% BSA in PBS. For off-target responses, plates were first coated with 2 μg/mL streptavidin (434302, Thermo Fisher Scientific), blocked with 2% BSA in PBS, and incubated with 2 μg/mL of HR1, C1, or V3 peptides diluted in blocking buffer. To detect antigen-specific IgG responses, dilutions of serum were added to the wells and incubated for 1.5 hours at 25°C. Plates were washed 3 times in PBS containing 0.2% Tween-20, then anti-mouse IgG secondary antibody conjugated to HRP (172–1011, Bio-Rad Laboratories), diluted 1:5000 in blocking buffer as per manufacturer instructions, was added to the wells. After 1 hour incubation, plates were again washed, developed with TMB, and stopped with sulfuric acid. Endpoint titers are reported as inverse dilutions where absorbance at 450 nm minus the reference absorbance at 540 nm equals 0.1.
Tavernier J., Tuypens T., Verhee A., Plaetinck G., Devos R., Van der Heyden J., Guisez Y, & Oefner C. (1995). Identification of receptor-binding domains on human interleukin 5 and design of an interleukin 5-derived receptor antagonist.. Proceedings of the National Academy of Sciences of the United States of America, 92(11), 5194-5198.
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9

Complement Binding ELISA Protocol

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High binding ELISA plates (07-200-37, Fisher Scientific) were coated with 1 µg/mL of labeled and blocked with 2% BSA in PBS. Serum was collected from naïve mice (41.1500.005, Sarstedt), diluted in PBS (3% v/v), and incubated in ELISA plates for 1 hour at 37°C. Complement binding was detected by biotinylated anti-C3 antibody (NB200-540B, Novus Biologicals) followed by streptavidin-HRP (3310-9-1000, Mabtech AB). The plates were developed with 1-Step Ultra TMB-ELISA Substrate Solution (34028, Thermo Fisher Scientific) and the reaction was stopped by adding with 2 N sulfuric acid (BDH7500-1).
Aung A., Cui A., Amini A.P., Bukenya M., Lee H., Cottrell C.A., Silva M., Kirkpatrick J.D., Gregory J.R., Amlashi P., Remba T., Froehle L.M., Xiao S., Abraham W., Adams J., Suh H., Huyett P., Kwon D.S., Hacohen N., Schief W.R., Bhatia S.N., & Irvine D.J. (2021). Spatially regulated protease activity in lymph nodes renders B cell follicles a sanctuary for retention of intact antigens.
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