Immulon 2 high binding 96-well plate (VWR) was coated with 500 ng/well of recombinant human PD-L1 (BioVision). After blocking plate with PBS-T containing 3% BSA, serially diluted media of A549-infected with 1,000 vp/cell of HDegfp or HDPD-L1 mini were added and incubated at 4°C for 24 hours. Serially diluted anti-human PD-L1 antibody starting from 10 μg/well (BioLegend) was used as a positive control. After washing plate with PBS-T, HRP-labeled anti-human IgG for PD-L1 mini-body detection or HRP labeled anti-mouse IgG (BioRad) for anti-human PD-L1 and isotype antibody detection were added and incubated at room temperature for 1 hour and then we developed the washed plate. Absorbance was measured using Tecan reader (TECAN).
Immulon 2 high binding 96 well plate
Manufactured by Avantor
Sourced in United States
The Immulon 2 high binding 96-well plate is a laboratory equipment designed for use in various immunoassay and binding studies. It features a high-binding surface that enhances the adsorption of proteins, peptides, and other biomolecules, making it suitable for a range of applications that require the immobilization of these substances on a solid support.
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2 protocols using immulon 2 high binding 96 well plate
1
PD-L1 Binding Assay Protocol
2
Minibody Binding ELISA for PD-L1, PD-1, and CTLA-4
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The minibody binding ELISA assay has been described previously [10 (link)]. Briefly, Immulon 2 high binding 96-well plate (VWR, Radnor, PA) was coated with 500 ng/well of recombinant human PD-L1, PD-1, or CTLA-4 (BioVision, Milpilas, CA, USA). After blocking plate with PBS-T containing 3% BSA, serially diluted media containing minibody or media from GFP-transfected cells were added and incubated at 4 °C for 24 h. Serially diluted IgGs (Biolegend, San Diego, CA, USA) were used as positive controls with isotype IgGs serving as negative controls. HRP-labeled anti-human IgG for minibody detection or HRP-labeled anti-mouse IgG (BioRad, Hercules, CA, USA) for anti-human antibody detection were added and incubated at room temperature for 1hr and then developed. Absorbance was measured using Tecan plate reader (TECAN, Männedorf, Switzerland).
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