About trypsin without phenol red - (Jan/28/2008 )
Please help
As i am doing a hormone withdrawal treatment of the MCF-7 and T-47D, after starvation, I need to to seed the cell for treatment.
During the seeding process, i need to trypsinize the cell. In the very begining I use Invitrogen 0.25% trypsin-EDTA with phenol red. However, one technician had said that phenol red is estrogenic and may affect the status of the cell.
I want to know would the effect of phenol red in this process affect much.
Now I am searching the Trypsin with no phenol red in Invitrogen, however, only 10X without EDTA is availiable.
Do the effect of trypsin without EDTA good enough? As I know if long time treatment of trypsin on cell is not good. Sometime after I use 0.25% trypsin-EDTA with phenol red, some white precipitate which I thought is dead cell after starvation... especially in the case of T-47D
Besides, I don't why after I treated the cell with E2 and tamoxifen citrate, the cell does not grow much faster or dead faster even in 1E-6M of E2 and 1E-5M of tamoxifen citrate.
Are my cell lines, MCF-7 and T47D, have problem...? 
As i am doing a hormone withdrawal treatment of the MCF-7 and T-47D, after starvation, I need to to seed the cell for treatment.
During the seeding process, i need to trypsinize the cell. In the very begining I use Invitrogen 0.25% trypsin-EDTA with phenol red. However, one technician had said that phenol red is estrogenic and may affect the status of the cell.
I want to know would the effect of phenol red in this process affect much.
Now I am searching the Trypsin with no phenol red in Invitrogen, however, only 10X without EDTA is availiable.
Do the effect of trypsin without EDTA good enough? As I know if long time treatment of trypsin on cell is not good. Sometime after I use 0.25% trypsin-EDTA with phenol red, some white precipitate which I thought is dead cell after starvation... especially in the case of T-47D
Besides, I don't why after I treated the cell with E2 and tamoxifen citrate, the cell does not grow much faster or dead faster even in 1E-6M of E2 and 1E-5M of tamoxifen citrate.
Are my cell lines, MCF-7 and T47D, have problem...?
Invitrogen offers f.i. TrypLE without phenol red; by the way, I do not a serious need for phenol red in trypsin/EDTA solution
What about using Cellstripper from Cellgro. It is non-enzymatic and does not contain phenol red.
You could make your own trypsin from powder?? And then add EDTA also if you wanted to??