Antibodies for detection of total serum are listed (Key Resource Table ). 96-well flat-bottom ELISA plates (Sarstedt) were coated with 75 μL per well of respective unlabeled coating antibody (diluted in PBS) 4°C overnight. Plates were blocked with PBS 1% BSA at room temperature for 1h. Plates were washed three times with PBS Tween20 and dH2O. Samples and standards were added to the wells and blank wells were included (PBS 1%BSA). Plates were incubated for 2–4h at 37°C before being washed three times. Secondary antibody, HRP (horseradish peroxidase) diluted in PBS 1%BSA was added and incubated at 37°C for 2h. The plates were then washed five times and 100 μL of OPD (o-phenylenediamine dihydrochloride) substrate solution (ThermoFisher Scientific) was added and plates incubated room temperature in the dark for 10–30 minutes to develop. OD values were read at 405nm using a plate reader (BMG Labtech) and the data was evaluated using MARS (BMG Labtech).
96 well flat bottom elisa plates
Manufactured by Sarstedt
The 96-well flat-bottom ELISA plates are a standard laboratory equipment used for enzyme-linked immunosorbent assay (ELISA) experiments. These plates have a flat-bottom design and 96 individual wells, allowing for the simultaneous testing of multiple samples or standards.
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2 protocols using 96 well flat bottom elisa plates
1
Serum Antibody Quantification ELISA
2
Serum Antibody Quantification ELISA
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Antibodies for detection of total serum are listed (Key Resource Table ). 96-well flat-bottom ELISA plates (Sarstedt) were coated with 75 μL per well of respective unlabeled coating antibody (diluted in PBS) 4°C overnight. Plates were blocked with PBS 1% BSA at room temperature for 1h. Plates were washed three times with PBS Tween20 and dH2O. Samples and standards were added to the wells and blank wells were included (PBS 1%BSA). Plates were incubated for 2–4h at 37°C before being washed three times. Secondary antibody, HRP (horseradish peroxidase) diluted in PBS 1%BSA was added and incubated at 37°C for 2h. The plates were then washed five times and 100 μL of OPD (o-phenylenediamine dihydrochloride) substrate solution (ThermoFisher Scientific) was added and plates incubated room temperature in the dark for 10–30 minutes to develop. OD values were read at 405nm using a plate reader (BMG Labtech) and the data was evaluated using MARS (BMG Labtech).
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