BioLux Gaussia Luciferase Assay Buffer can be stored at 4°C.
BioLux Gaussia Luciferase Substrate must be tightly capped and stored at -20°C.
Usage notes:
Because of the stability of GLuc, the activity measured in the growth media of GLuc-expressing culture reflects the protein that has accumulated up to the time of sampling.
Equilibration of the GLuc assay solution is not necessary.
After adding the GLuc assay solution to the sample, we recommend a delay time of 5–15 seconds before taking a measurement. Keeping the delay time consistent across experiments will ensure reproducibility.
Use the prepared assay solution within 24 hours. The unused portion of the assay solution should be tightly capped and stored at -20°C. It should be completely thawed (in the dark) to room temperature before use.
The linear range of the luminometer used for the assay must be established. This is easily done by assaying serial dilutions of a sample. In addition, the assay solution itself as well as the conditioned media (i.e. growth media from untransfected cells) should be included to establish the background in the assay.
If excess activity for the instrument range is found, the sample should be diluted in PBS or 10% serum-containing media. The integration time can also be reduced (e.g. 2 seconds instead of 5 seconds).
When assaying the serial dilutions of a sample, it is best to assay the most diluted samples first & the most concentrated samples last. This will help to minimize false readings, i.e. cross talk effect in which signals from samples of high RLU cross into the next sample. The cross-talk effect seems to be more pronounced when white or black plates with clear-bottoms are used.