Maxisorb plate

Manufactured by Thermo Fisher Scientific

Sourced in Denmark, United States, Germany

Maxisorb plates are a type of laboratory equipment used for various applications. They are designed to provide a high surface area for the efficient adsorption and immobilization of substances. The core function of Maxisorb plates is to facilitate the process of adsorption, which is crucial in various scientific and analytical procedures.

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Lab products found in correlation

Hrp conjugated goat anti llama igg, by Fortis Life Sciences (4 mentions) Clariostar plate reader, by BMG Labtech (3 mentions) Type 1 collagen from rat tail, by BD (3 mentions) Igg2c, by Thermo Fisher Scientific (3 mentions) Tmb single substrate solution, by Merck Group (2 mentions) Goat anti mouse hrp conjugate, by Merck Group (2 mentions) Multiskan fc, by Thermo Fisher Scientific (2 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (2 mentions) Human fibronectin, by Serva Electrophoresis (2 mentions) Gelatin, by Merck Group (2 mentions) O phenylenediamine opd, by Merck Group (2 mentions) Streptavidin hrp, by BD (2 mentions) β galactosidase, by Merck Group (2 mentions) Horseradish peroxidase hrp coupled streptavidin, by Jackson ImmunoResearch (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Elispot kit, by Mabtech (1 mentions) Anti cd28, by BD (1 mentions) Anti ha antibody, by Abcam (1 mentions) P0260, by Agilent Technologies (1 mentions) Sureblue tmb substrate, by LGC (1 mentions)

71 protocols using maxisorb plate

Quantifying Total IgE and IgE-secreting Cells

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For total IgE ELISA, MaxiSorb plates (ThermoFisher Scientific) were coated with 0.5 μg/ml anti-human IgE (555894, BD Pharmingen) in carbonate-bicarbonate buffer overnight at 4°C. Coated wells were blocked with 5% skim milk in PBS for 2 hours at room temperature, followed by 3 washes (1x PBS and 0.05% Tween 20). Cell-free supernatant samples and a serial dilution of purified human IgE (401152, Calbiochem) were incubated overnight at 4°C. Wells were washed 3 times and incubated with 1 μg/ml biotinylated anti-human IgE (A18803, Invitrogen) in 1% skim milk for 2 hours at room temperature. Subsequently, wells were washed 3 times and incubated with alkaline-phosphatase streptavidin for 1 hour at room temperature. Plates were developed with p-nitrophenyl phosphate and the reaction was stopped with 2N NaOH. Optical density was measured at 405 nM (Multiskan™FC, Thermo Scientific).
A commercially available ELISPOT kit (3810–2H, Mabtech) was used for the detection of IgE-secreting cells. On day 11 of culture, samples were plated in duplicate at 4 × 10⁶ cells/mL. Plates were imaged with ImmunoSpot® S6 Analyzer and spots were counted independently by 2 blinded investigators.

Jiménez-Saiz R., Ellenbogen Y., Bruton K., Spill P., Sommer D.D., Lima H., Waserman S., Patil S.U., Shreffler W.G, & Jordana M. (2019). Human BCR analysis of single-sorted, putative IgE+ memory B cells in food allergy. The Journal of allergy and clinical immunology, 144(1), 336-339.e6.

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PBMC and T-cell Activation with Anti-CD3 and Anti-CD28

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1 × 10⁵ PBMCs or T cells were incubated in 96-well Maxi-Sorb plates (ThermoFisher) with 1 μg anti-CD3 (OKT3, purified from hybridoma supernatant) and, where indicated, 100 nM UL11Fc or Fc control preadsorbed onto the surface. For the T cells, 2 μg/mL of anti-CD28 (Clone L293, BD Pharmingen) were added to the medium. To generate the CMV-specific responses, 100 μg of HCMV-infected cell lysate (antibodies-online GmbH) was adsorbed onto the plate, and anti-CD28 (2 μg/mL) was added to the medium.

Osanyinlusi S.A., Zischke J., Jacobs R., Weissinger E.M., Schulz T.F, & Kay-Fedorov P.C. (2022). Human Cytomegalovirus pUL11, a CD45 Ligand, Disrupts CD4 T Cell Control of Viral Spread in Epithelial Cells. mBio, 13(6), e02946-22.

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Serum IgE and IgG4 Binding Assay

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Shrimp extract of Pm and Mr was prepared as previous described.8 (link) To measure serum IgE and IgG4 to allergen in the Pm and Mr extract by indirect binding ELISA, shrimp extract was diluted in phosphate-buffered saline (PBS) and coated individually at 500 ng per well on 96-well Maxisorb plates (Nunc™, Thermo Fisher Scientific, Waltham, MA, USA) and incubated at 4°C overnight. Moreover, to measure serum IgE and IgG4 binding to recombinant TM, AK and MLC, each recombinant allergen was also diluted in PBS and coated individually at 500 ng per well on 96-well Maxisorb plates (Nunc™, Thermo Fisher Scientific, Waltham, MA, USA) and incubated at 4°C overnight. The coated 96-well plate was washed with PBS-A (PBS+3% non-fat dried milk). Sera of patients and controls were diluted at 8:1 in PBS-A. Diluted sera were incubated with coated allergens in PBS-A for 2 hours at room temperature. The plate was washed before either diluted HRP-labelled goat IgG anti-human IgE or diluted HRP-labelled rabbit IgG anti-human IgG4 antibodies was added into the designated wells. Substrate (3,3′,5,5′-tetramethylbenzidine or TMB, Thermo Fisher Scientific) was added to each well before absorbance at OD_{650 nm} was measured.

Piboonpocanun S., Ittiporn S., Ubonsri P., Wangtan A., Pacharn P., Visitsunthorn N, & Jirapongsananuruk O. (2022). Defining Biomarkers to Predict Natural Resolution in Shrimp Allergy. Allergy, Asthma & Immunology Research, 14(2), 210-219.

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Screening DARPin Binding to CD8

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To screen the output of the DARPin selection for binding to CD8, an ELISA with recombinant CD8-Fc_Biotin protein was conducted as previously described.³¹ (link) In brief, 96-well Maxisorb plates (Thermo Fisher Scientific) were coated with 100 mL 20 mM Neutravidin solution in TBS for 1 h at room temperature. After washing, unspecific binding was blocked with TBS-T supplemented with 0.5% BSA for 1 h at room temperature. To immobilize the target protein, wells were coated with 10–20 nM of biotinylated CD8αα-Fc, CD8αβ-Fc, or Fc-only protein solution in blocking buffer. After washing, wells were incubated with diluted DARPin crude lysate preparations for 1 h at room temperature. To quantify the amount of bound DARPin proteins, wells were subsequently incubated with anti-HA antibody (1:5,000, Abcam, Cambridge, UK, clone 16B12, ab130275) and a horseradish peroxidase (HRP)-conjugated rabbit anti-mouse antibody (1:1,000; P0260, Agilent, Santa Clara, CA, USA) for 1 h at room temperature each. The plates were washed three times with TBS-T before and after each antibody incubation step. The bound antibodies were detected using SureBlue TMB substrate (SeraCare, Milford, MA, USA) and terminated after 5–10 min using 1N H₂SO₄. The reaction product was quantified in a microtiter plate reader at 450 nm.

Michels A., Frank A.M., Günther D.M., Mataei M., Börner K., Grimm D., Hartmann J, & Buchholz C.J. (2021). Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8. Molecular Therapy. Methods & Clinical Development, 23, 334-347.

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ELISA Assay for NTHi Antibody Titration

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ELISA assays were carried out using standard protocols⁶⁸ in 96-well Maxisorb plates (NUNC; Thermo Scientific). Cells were diluted to a 0D₆₀₀ of 0.2 (2 × 10⁸ c.f.u. ml⁻¹), with 50 μl added per well. All the strains were assayed in triplicate. Primary antibody AD6 against HMW^{34 (link)} was used at a starting concentration of 1:200 (NTHi strains C486 and 477) or 1:10,000 (NTHi strain 723) and serially diluted two-fold in 1 × PBS pH 7.9. Secondary antibody (goat anti-mouse HRP conjugate; Sigma-Aldrich) was used at a concentration of 1:10,000. Antibody was detected using TMB single-substrate solution as recommended by the manufacturer (Sigma-Aldrich). The data were plotted as antibody dilution (x axis) versus absorbance at 450 nm (y axis), and data from specific titres in the linear range of the response curve picked for statistical analysis.

Atack J.M., Srikhanta Y.N., Fox K.L., Jurcisek J.A., Brockman K.L., Clark T.A., Boitano M., Power P.M., Jen F.E., McEwan A.G., Grimmond S.M., Smith A.L., Barenkamp S.J., Korlach J., Bakaletz L.O, & Jennings M.P. (2015). A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae. Nature Communications, 6, 7828.

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Enzyme-Linked Immunosorbent Assay for VHH Binding

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After overnight immobilization at 4 °C with the WT46 or WT52 (2 μg/mL) onto maxisorb plates (Thermo Scientific), the wells were washed with PBS (0.005% v/v) -Tween-80 and blocked with milk protein in PBS (4% w/v, Marvel). A serial dilution of the VHHs anti-WT46 and anti-WT52 was then performed (0–1000 nM) and incubates 1h at room temperature before addition of an anti-Flag Tag Ab (M2, 1/5000, Sigma-Aldrich) 1h at room temperature. After a washing step, a secondary donkey anti-mouse Ab coupled to a horseradish peroxidase (1/5000, Jackson ImmunoResearch) was incubated 1h at room temperature. After a last washing step, o-Phenylenediamine dihydrochloride substrate was added (1/1000, Sigma-Aldrich), the reaction was stopped with 1 M sulfuric acid and the optical density was measured at 490 nm.

Godar M., Morello V., Sadi A., Hultberg A., De Jonge N., Basilico C., Hanssens V., Saunders M., Lambrecht B.N., El Khattabi M., de Haard H., Michieli P, & Blanchetot C. (2016). Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy. Scientific Reports, 6, 31621.

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ELISA Assay for HMW34 Antibody

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ELISA assays were carried out using standard protocols68 in 96-well Maxisorb plates (NUNC; Thermo Scientific). Cells were diluted to a 0D₆₀₀ of 0.2 (2 × 10⁸ c.f.u. ml⁻¹), with 50 μl added per well. All the strains were assayed in triplicate. Primary antibody AD6 against HMW34 (link) was used at a starting concentration of 1:200 (NTHi strains C486 and 477) or 1:10,000 (NTHi strain 723) and serially diluted two-fold in 1 × PBS pH 7.9. Secondary antibody (goat anti-mouse HRP conjugate; Sigma-Aldrich) was used at a concentration of 1:10,000. Antibody was detected using TMB single-substrate solution as recommended by the manufacturer (Sigma-Aldrich). The data were plotted as antibody dilution (x axis) versus absorbance at 450 nm (y axis), and data from specific titres in the linear range of the response curve picked for statistical analysis.

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ELISA for Tau Protein Detection

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Maxisorb ® plates (Thermo Scientific, USA) were coated with 5µg/ml 1G2 antibody in 0.05 mol/l Na2CO3/NaHCO3, pH 9.6 with 120 µl per well at 4-8°C overnight. Coated plates were aspirated and blocked using 50 mM Tris buffer pH 7.5 containing 0.15 M NaCl and 0.05% Tween 20 (washing buffer) containing 3 % BSA fraction V (Serva, Heidelberg, Germany). Lyophilized standards were designed using different concentrations of Tau2N4R (rPeptide) as well as controls and negative controls by means of phosphate buffered saline buffer containing BSA and stabilizers. The assay was optimised studying different incubation protocols, and 3 h room temperature on orbital shaker appeared appropriate for antigen binding in the first step.
Washing was performed 5 times using 50 mM Tris buffer pH 7.5 containing 0.15 M NaCl and 0.05% Tween 20 and followed by 90 min incubation with HRP conjugated detection antibody 7E5 (Supplementary Material) at room temperature. This antibody is described as anti-human Tau antibody that binds Tau2N4R between amino acids 156-165. After 5 additional washing steps, the plate was developed with TMB substrate solution for 30 min in the dark followed by termination with 1.5 M H2SO4. Optical density (OD) was measured at 450nm as well as 450/620 nm.

Lewczuk P., Lelental N., Lachmann I., Holzer M., Flach K., Brandner S., Brandner S., Engelborghs S., Teunissen C.E., Zetterberg H., Molinuevo J.L., Mroczko B., Blennow K., Popp J., Parnetti L., Chiasserini D., Perret-Liaudet A., Spitzer P., Maler J.M, & Kornhuber J. (2017). Non-Phosphorylated Tau as a Potential Biomarker of Alzheimer's Disease: Analytical and Diagnostic Characterization. Journal of Alzheimer's disease : JAD, 55(1).

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ELISA for Antibody Titer Quantification

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Enzyme-Linked Immunosorbent Assay (ELISA) was used to determine the antibody titers. Maxisorb plates (Nunc) were coated with 5 µg of recombinant protein and were blocked with 5% skim milk in PBST (PBS plus 0.05% Tween 20). The washed plates were incubated with serially diluted serum samples at 37°C for 30 min. The plates were washed four times with PBST. Then, 100 µl of 1 in 2000 dilution of Horseradish Peroxidase (HRP)-conjugated goat anti-mouse IgG were added to each well plate. After that, the plate was incubated at 37°C for 30 min and was then washed four times with PBST. Meanwhile, the plates were incubated with HRP-conjugated anti-mouse monoclonal IgG1 or IgG2a under the same conditions. Then, 100 µl of the substrate solution containing 3 mg O-Phenylene Diamine (OPD) were added to each well plate. Finally, the reaction was stopped with 2.5 M H₂SO₄ and the absorbance was read at A₄₉₅nm on a microplate reader.

Parvane M., Nazarian S., Kordbache E., Fathi J., Minae M.E, & Ramezani M.R. (2022). Evaluation of PLGA-Encapsulated Recombinant GroEL of S. typhi immune Responses Against Enterohaemorrhagic and Enteropathogenic Escherichia coli. Avicenna Journal of Medical Biotechnology, 14(4), 294-302.

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Recombinant CDPC1 ELISA Protocol

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Enzyme-linked immunosorbent assays (ELISAs) were performed by immobilising 50 μL recombinant CDPC1 per well, typically at 1–5 μg/ml in PBS, on 96-well Maxisorb plates (Nunc) overnight at 4 °C. Bovine serum albumin (Sigma) at the same concentration was added as a negative control antigen to wells on the same plate. The antigen plates were washed three times with PBS and blocked in 3 % non-fat milk powder in PBS, then 50 μL/well antibodies in blocking solution were added and incubated at room temperature for at least 1 h. The plates were washed three times with PBS-Tween, then incubated with 50 μL of appropriate secondary antibody-peroxidase conjugates (goat anti-human-Fc-peroxidase conjugate, Sigma cat # A0170, at 1:10,000 dilution was used for human IgGs and DARpin-Fcs) in 3 % non-fat milk in PBS-Tween for at least 30 min at room temperature, washed again three times in PBS-Tween, and developed for 2–10 min using 50 μL 3,3′,5,5′-Tetramethylbenzidine (TMB) substrate. The reaction was quenched with 50 μL 0.5 M H₂SO₄, then the absorbance measured at 450 nm on an Envision microplate reader.

Sandercock A.M., Rust S., Guillard S., Sachsenmeier K.F., Holoweckyj N., Hay C., Flynn M., Huang Q., Yan K., Herpers B., Price L.S., Soden J., Freeth J., Jermutus L., Hollingsworth R, & Minter R. (2015). Identification of anti-tumour biologics using primary tumour models, 3-D phenotypic screening and image-based multi-parametric profiling. Molecular Cancer, 14, 147.

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