Nitrocef disks

Reporter gene assay β-lactamase substrates - HEK 293 & HEK 293T cells

Experiment
Reporter gene assay β-lactamase substrates - HEK 293 & HEK 293T cells
Product
Nitrocef disks from Hardy Diagnostics
Manufacturer
Hardy Diagnostics

Protocol tips

Protocol tips
Allow the Nitrocef Disk™ to equilibrate to room temperature.

Publication protocol

For the βlac assay, 1 × 105 cells were plated into individual wells of a poly‐d‐lysine‐coated 48‐well plate. The assay was performed 24 h after plating. Nitrocefin was first diluted to a final concentration of 100 μM in PBS. After drug treatment, the cells were washed once with PBS and, after removal of the PBS wash, 200 μL of the nitrocefin solution was added to the each well. Immediately after the addition of the nitrocefin solution, absorbance for each well was read once every minute for 30 min at 486 nm using the EPOCH microplate spectrophotometer (Biotek). The rate of reaction (slope of the curve in the linear range) was taken as the readout for this assay (see Figure S4 for an example).

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Papers

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Paper title
A novel assay for measurement of membrane-protein surface expression using a β-lactamase.
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Manufacturer protocol

Download the product protocol from Hardy Diagnostics for Nitrocef disks below.

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