The nAb titers against HAdV4 and HAdV7 were determined using neutralization assays based on HAdV4-SEAP and HAdV7-SEAP, respectively. In brief, HEK293 cells were seeded into 96-well plates at 3 × 104 cells per well. One day later, HAdV4-SEAP or HAdV7-SEAP at 4 × 106 vp per well (∼100 vp/cell) were incubated for 1 h at 37°C either alone (virus infection alone) or together with serial dilutions of human sera or mouse antisera. Subsequently, each sample was added to the 96-well plates and incubated for 24 h at 37°C. Finally, the supernatants were harvested and the SEAP activity was assessed using the chemiluminescent substrate CSPD according to the manufacturer’s protocols (Thermo Fisher Scientific, Waltham, MA, United States). The relative light units (RLU) were recorded using a luminometer (MlX Microtiter, Dynex Technologies, Inc., United States) and the titers were calculated as the dilutions that inhibited 50% of RLU values.
Mlx microtiter
Manufactured by Dynex
Sourced in United States
The MlX Microtiter is a laboratory instrument used for conducting microplate-based assays. It is designed to facilitate the measurement and analysis of various samples in a multi-well format. The core function of the MlX Microtiter is to provide a platform for performing a wide range of analytical techniques, such as absorbance, fluorescence, and luminescence measurements, on small-volume samples.
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Lab products found in correlation
2 protocols using mlx microtiter
1
Neutralization Assay for Adenovirus Antibodies
2
Serum nAb Titers Against HAdV26 and HAdV35
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The serum neutralizing antibody (nAb) titers against HAdV26 and HAdV35 were detected by MN assays as previously established in our lab. In brief, the HEK293 cells were seeded into 96-well plates at 3 × 104 cells per well and cultured for 36 h. Human serums were serially three-fold diluted in DMEM from 1:18 to 1:18,432, and incubated with HAdV26-SEAP or HAdV35-SEAP at 5 × 106 vp per well for 1 h at 37 °C. After that, the mixtures were added to the HEK293 cells and incubated at 37 °C for 24 h. At last, the supernatants were harvested to detect SEAP activity with the Phospha-Light System (Thermo Fisher Scientific, Cat: T1016, Massachusetts, USA) according to the manufacturer's instructions. Relative light units (RLU) were recorded by a luminometer (MLX Microtiter®, Dynex Technologies, Inc., USA). nAb titers from duplicated wells were calculated as the dilutions that inhibited 50% RLU values.
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