Synthesis of PHP-APED

Synthesis

9/2/2014

1,2-Bis(3-aminopropylamino)ethane, APED
 
http://www.sigmaaldrich.com/catalog/product/aldrich/239399
239399-100ML, $29.30

	DBP	DAP	APED	MeOH
D1	61 mg	12.5 mg	5 ul	75 ul
D2	61	12.5	10	75
D3	61	12.5	15	75
D4	61	12.5	20	75
D5	61	12.5	25	75
D6	61	12.5	30	75

366 mg DBP + 75 mg DAP+ 150 ul MeOH-->335 ul

Divide into 6 vials

+ 5 to 30 ul APED

+ 75 ul MeOH

50 ^oC react o/n

then 60 ^oC react 3 hours.

Purify with 0.5 ml isopropanol, 2X

Dissolve in 500 ul H2O

Transfection

9/4/14 Transfection: dilute 1:10, 1:30, 1:50, 1:75, use 3 ul + 1 ug DNA to transfect 293T cells in 24 well plate

9/5/14 Morning

9/5/14 Afternoon

9/6/14 Morning

9/7/14

9/8/14

Comparison of TREN1, PHP-TREN, PHP-s-s-TREN and PHP-APED

TREN1, PHP-TREN (A2, A3, B1): see this page: Synthesis_of_PHP-TREN

PHP-TREN (C1): Synthesis_of_PHP-s-s-TREN

PHP-APED (D6)

Transfection

9/5/14

Dilution: 1:10, 10:30, 1:50, 1:75

DNA 1 ug

293 cells in 24 wells

Result

9/6/14:

Dead cells

	TREN1	C1	D6	A2	A3	B1
1:10	99%	95%	50%	25%	40%	20%
1:30	99%	10%	2%	15%	15%	1%
1:50	95%	30%	20%	15%	15%	1%
1:75	80%	5%	40%	15%	2%	1%

9/6/14 Morning

9/6/14 Afternoon

9/7/2014

9/8/14