Ten micrograms of whole-cell lysates of DBT cells infected with cl-2 or JHM-X at an MOI of 0.002 or mock infected were coated on microtiter plates (SPL, Korea) at 4 °C overnight. Then, the plates coated with lysates were incubated with the monoclonal antibody 10 G diluted at 1:5000 in 5% BSA solution at 4 °C overnight after blocking with 5% BSA for 2 h at room temperature. The plates were washed with PBSt three times for 5 min each time and incubated with HRP-conjugated anti-mouse IgG (CSB-PA644737, Cusabio, USA) at room temperature for two hours. Following 3 washes with PBSt, the plates were incubated with TMB substrate (Komabiotech, Korea) in the dark for 5–10 min, followed by incubation with stop solution (Komabiotech, Korea). Absorbance (optical density) values were measured at a wavelength of 450 nm using a VICTOR® Nivo™ Multimode Plate Reader (PerkinElmer).
Microtiter plates
Manufactured by SPL Life Sciences
Microtiter plates are a type of laboratory equipment used to perform various assays and experiments. They are flat plates, typically made of plastic, with multiple small wells arranged in a grid pattern. These wells are designed to hold small volumes of liquid samples, enabling the simultaneous analysis of multiple samples or the performance of multiple tests on a single plate. Microtiter plates are widely used in fields such as biochemistry, cell biology, and molecular biology for applications like enzyme-linked immunosorbent assays (ELISAs), cell-based assays, and high-throughput screening.
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Lab products found in correlation
2 protocols using microtiter plates
1
Quantification of SARS-CoV-2 Spike Protein Levels
2
Rat Brain NMU Content Quantification
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An indirect sandwich ELISA was developed to measure the rat brain NMU content. Briefly, brain tissue homogenated in phosphate-buffered saline (PBS, pH 7.2) and centrifuged at 13,000 rpm for 10 min. The supernatant used for ELISA assays. 100 μl of the NMU- goat polyclonal antibody (Santa-cruz Biotechnology), diluted to 1:50 in a coating buffer (100 mM carbonate/bicarbonate buffer, pH 9.6), was used to coat the wells of microtiter plates (SPL Life Sciences) except for three wells as control wells and incubated at 4 °C overnight. Then the wells were washed with PBS containing 0.05% Tween 20 (PBST) for three times and blocked with 1% bovine serum albumin (BSA) in PBS at 37 °C for 90 min to diminish nonspecific binding. Afterward, they were incubated in a 1:100 dilution of primary anti-NMU in PBST at 37 °C for 2 hr, washed for 3 times in PBST and incubated for 2 hr with a donkey anti-goat peroxidase conjugate secondary Ab diluted 1/10000 in PBST (Santa Cruz Biotechnology).
Washing was done with PBST for 4 times and 100 μl of TMB (3,3V, 5,5V-tetramethylbenzidine) as a substrate was added. The reaction was quenched using 0.15 M H2SO4 (100 μl) after appropriate development time and the optical density (OD) was determined with an ELISA reader (DRG Elisa-Mat 2000) at 450 nm.
Washing was done with PBST for 4 times and 100 μl of TMB (3,3V, 5,5V-tetramethylbenzidine) as a substrate was added. The reaction was quenched using 0.15 M H2SO4 (100 μl) after appropriate development time and the optical density (OD) was determined with an ELISA reader (DRG Elisa-Mat 2000) at 450 nm.
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