How long does it take for GFP to degrade?
The half-life of unmodified GFP is approximately 26 h;8 thus, it takes several days for the passively transferred protein to degrade leading to an overestimation of transduction achieved at early time points.
How long does GFP last in a cell?
The half-life of the green fluorescent protein (GFP) was determined biochemically in cultured mouse LA-9 cells. The wild-type protein was found to be stable with a half-life of ~26 h, but could be destabilized by the addition of putative proteolytic signal sequences derived from proteins with shorter half-lives.
Does GFP degrade?
We found that GFP degradation rates were independent of the initial intensity of GFP fluorescence within cells. This result indicates that higher GFP abundance in some cells is likely due to higher rates of gene expression, because it is not due to systematically lower rates of protein degradation.
How long does it take for GFP to be expressed?
GFP expression was noticeable in cells within 4 h of transfection. In nine separate transfections, approximately 20% of the transfected cells expressed GFP with a mean fluorescence 40-50x that of control cells (15 fluorescent units [FU] vs. 0.3 FU) during the first five days after transfection.
What is d2EGFP?
Our data suggest that d2EGFP is a useful indicator of proteasome inhibition. Therefore, stable expression of d2EGFP in mammalian cells is potentially useful for high-throughput screening of cDNAs or pharmaceutical drugs that repress proteasome functions in vivo.
How long after transfection is GFP?
Does GFP fade over time?
Studies have shown that GFP expression can deteriorate over time as GFP tagged cells are prone to death. Therefore, the cells that were originally marked with GFP do not survive and cannot be accurately traced over time.
What is mEGFP?
mEGFP is a basic (constitutively fluorescent) green fluorescent protein published in 2002, derived from Aequorea victoria. It has moderate acid sensitivity.
How long does transfection expression take?
Post-transfection Incubation Time In general, maximal protein expression occurs 48 hours post-transfection. The time point for optimal gene expression can be determined by varying post-transfection incubation times from 4 to 72 hours.