Q: Will this kit detect EGFP?
A: This kit will detect fluorescence from EGFP. The recombinant GFP standard used in the kit is also EGFP.
Q: What is the protocol for lysing cells?
A: Here is the protocol we recommend for lysing cells:
Wash cells once with cold PBS, then add cold 1X Assay/Lysis Buffer and incubate 5-10min at 4C. Collect lysate (supernatant) by centrifuging 10,000g for 5 minutes at 4C.
Q: Can lysates be stored prior to using the assay?
A: GFP is very stable molecule and as long as your lysis buffer contains proteinase inhibitors the lysates can be stored at -20C for three months or at -80C for one year.
Q: Should I add my samples at a specific protein concentration?
A: We don’t recommend a specific total protein concentration because detection is based on the levels of GFP present in the sample, and this will vary between samples. The limit of detection with AKR-120 is 156ng/ml GFP.