Journal: Micromachines

Article Title: A Nitrocellulose Paper-Based Multi-Well Plate for Point-of-Care ELISA

doi: 10.3390/mi13122232

Figure Lengend Snippet: The schematic representation of the ( a ) direct ELISA of rabbit IgG and ( b ) the sandwich ELISA of sperm protein SP-10 on the NC paper multi-well plate.

Article Snippet: The sandwich ELISA protocol is described as follows: The sandwich ELISA of SP-10 starts with the addition of 3 μL of the capture antibody solution (mouse anti-ACRV1 monoclonal antibody in CBS solution, cat #11789-MM01, Sino Biological) with a concentration of 100 μg/mL, onto the NC paper well, followed by the overnight incubation in a 4 °C fridge for the sufficient antibody immobilization, Then, after taking the blocked plate out of the fridge, each NC paper well was pipetted with 10 μL of the wash buffer (0.05% Tween20 in TBS, pH 7.2–7.4) and blotted with an absorbent pad, three times, to remove the unbound antibodies, Next, 3 μL of the blocking buffer (2% BSA in wash buffer) was added to each well ( (b-i)), followed by a 10-min incubation and washing step with 10 μL wash buffer, three times, again to remove the unbound blocking reagents ( (b-ii)), Next, 3 μL of the SP-10 protein solution (cat #10227-H08B, Sino Biological) was added to the blocked NC paper well and incubated for 10 min, for the SP-10 protein to be captured by the immobilized capture antibody probe ( (b-iii)).

Techniques: Direct ELISA, Sandwich ELISA

Journal: Micromachines

Article Title: A Nitrocellulose Paper-Based Multi-Well Plate for Point-of-Care ELISA

doi: 10.3390/mi13122232

Figure Lengend Snippet: The comparison between the SP-10 NC-paper-based ELISA and the standard plate-based ELISA for the SP-10 detection.

Article Snippet: The sandwich ELISA protocol is described as follows: The sandwich ELISA of SP-10 starts with the addition of 3 μL of the capture antibody solution (mouse anti-ACRV1 monoclonal antibody in CBS solution, cat #11789-MM01, Sino Biological) with a concentration of 100 μg/mL, onto the NC paper well, followed by the overnight incubation in a 4 °C fridge for the sufficient antibody immobilization, Then, after taking the blocked plate out of the fridge, each NC paper well was pipetted with 10 μL of the wash buffer (0.05% Tween20 in TBS, pH 7.2–7.4) and blotted with an absorbent pad, three times, to remove the unbound antibodies, Next, 3 μL of the blocking buffer (2% BSA in wash buffer) was added to each well ( (b-i)), followed by a 10-min incubation and washing step with 10 μL wash buffer, three times, again to remove the unbound blocking reagents ( (b-ii)), Next, 3 μL of the SP-10 protein solution (cat #10227-H08B, Sino Biological) was added to the blocked NC paper well and incubated for 10 min, for the SP-10 protein to be captured by the immobilized capture antibody probe ( (b-iii)).

Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Biomarker Assay