Microlon high binding

Manufactured by Greiner

Sourced in Germany

Microlon High binding is a laboratory equipment product designed for high-binding applications. Its core function is to provide a high-adsorption surface for various biomolecules and analytes during laboratory procedures.

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Lab products found in correlation

Granzyme b, by Mabtech (1 mentions) Biotinylated lectin, by Vector Laboratories (1 mentions) Europium conjugated streptavidin, by PerkinElmer (1 mentions) Carbohydrate free blocking solution, by Vector Laboratories (1 mentions)

Close spelling variants of this product

Microlon high-affinity binding plates (2 citations)

3 protocols using microlon high binding

Quantifying Cytokine Secretion in Activated T-Cells

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Cytokine secretion of antigen-specific activated T-cells was analyzed by ELISA. Commercial ELISA kits were used to measure the secreted cytokines Granzyme B and IL-6 (both from Mabtech AB, Nacka, Sweden), according to the manufacturer´s instructions. Briefly, a monoclonal antibody specific for Granzyme B or IL-6 was pre-coated onto a microplate (Microlon High binding, Greiner Bio One, Frickenhausen, Germany). Standards and samples were added into the wells, and any cytokine present was bound by the immobilized antibody. After washing to remove the unbound substances, an enzyme-linked monoclonal antibody specific for Granzyme B or IL-6 was added to the wells. Following washing to remove any unbound antibody-enzyme reagent, a substrate solution was added to the wells, leading to different coloring in proportion to the amount of Granzyme B or IL-6. The intensity of the color was analyzed with an automated plate reader (TECAN, Austria Gesellschaft, Grödig, Austria) using Magellan Software V2.22 at OD 450 nm.

Neuber B., Dai J., Waraich W.A., Awwad M.H., Engelhardt M., Schmitt M., Medenhoff S., Witzens-Harig M., Ho A.D., Goldschmidt H, & Hundemer M. (2017). Lenalidomide overcomes the immunosuppression of regulatory CD8+CD28− T-cells. Oncotarget, 8(58), 98200-98214.

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Lectin ELISA for Purified IgG

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Lectin ELISA was performed on purified IgG, as has been published with plasma samples [6 (link)]. Briefly, purified IgG was diluted to 1 mg/mL in carbonate coating buffer (100 mM NaHCO₃, 30 mM NaCO₃, pH 9.5), pipetted into a 96-well high-binding ELISA plate (Microlon High Binding; Greiner BioOne), and incubated overnight at 4 °C. The plate was blocked with carbohydrate free blocking solution (Vector Labs) for 1 hour at room temperature. Biotinylated lectins (Vector Labs) were diluted to 1 μg/mL in carbohydrate free blocking solution and incubated on the plate for 1 hour at room temperature. Signal was detected using europium-conjugated streptavidin (Perkin Elmer) and time-resolved fluorescence as measured in a Victor V3 1420 multilabel plate reader.

Glendenning L.M., Zhou J.Y., Reynero K.M, & Cobb B.A. (2022). Divergent Golgi Trafficking Limits B cell-Mediated IgG Sialylation. Journal of leukocyte biology, 112(6), 1555-1566.

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Multiplex ELISA for Autoantibody Detection

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Microtiter plates (MICROLON High Binding; Greiner) were coated with 100 ng/well EC26-2A4 or 3S peptide in PBS in duplicates at 4°C overnight. For the biotinylated EC26-2A4ΔM peptide, we used 10 ng/well and precoated streptavidin plates (200 ng/well). After three washing steps with PBST (PBS/0.05% Tween-20), plates were blocked with 5% (w/v) skimmed MPBST for 2 h at RT and subsequently incubated with patient plasma (diluted 1:100) or mouse sera (1:100–1:312,500 in fivefold serial dilutions) for 2 h at RT. Plates were washed thrice with PBST and incubated with HRP-conjugated anti-human or anti-mouse IgG antibodies, respectively, for 1 h at RT. After five washing steps, the plates were developed using 100 μl/well SureBlue TMB substrate (KPL, Inc.), stopped with 100 μl 1 N HCl, and read at 450 and 650 nm as a reference.

Ringel O., Müller K., Koch J., Brill B., Wolf T., Stephan C., Vieillard V., Debré P, & Dietrich U. (2018). Optimization of the EC26-2A4 Epitope in the gp41 Membrane Proximal External Region Targeted by Neutralizing Antibodies from an Elite Controller. AIDS Research and Human Retroviruses, 34(4), 365-374.

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