Use CPP to Promote Transduction
Use CPP to promote Transduction of Tat-Cre and Tat-Flp
1. Transfect 293T cells with reporters,
6 wells plate, 3 ug DNA each well
Cre: 2181 pCMV-LoxP-kozak-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Flp: 2013 pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP
2. Split cells next day into 12 or 24 wells plate
3. Add 0.5 μM Tat-Cre or Tat-Flp
and add the following peptides at 5 μM, make sure there are negative controls (No tat-Cre/Tat-Flp, No peptide)
Use this page to calculate MW to concentration
http://www.molbiol.ru/eng/scripts/01_04.html
=============
R9-HA
RRRRRRRRRGLFEAIEGFIENGWEGMIDGWYGC
MW: 4100
1 mg-->243.9 nmole, 1 mg in 1 ml--> 243.9 μM
=============
Tat-HA2
RRRQRRKKRGGDIMGEWGNEIFGAIAGFLGC
MW: 3536
1 mg-->285.7 nmole, 1 mg in 1 ml--> μM
=============
HA2-R9
GDIMGEWGNEIFGAIAGFLGGGRRRRRRRRRGC
MW: 3735
1 mg-->270 nmole, 1 mg in 1 ml--> 270 μM
=============
Tat-IFN7
RRRQRRKKRGLFEAIEGFIENGWEGMIDGWYGC
MW: 4016
1 mg-->250 nmole, 1 mg in 1 ml--> 250μM
4. Incubate o/n and take a picture of cells. Compare transduction efficiency.