Paul.xu: moved Synthesis EDA-CBA-PEI to Synthesis 2A1E-CBA-PEI

2011-11-17T20:56:30Z

moved Synthesis EDA-CBA-PEI to Synthesis 2A1E-CBA-PEI

New page

[[Image:Synthesis SS-PBAA-PEI.jpg|600px|Synthesis SS-PBAA-PEI.jpg]] 

 

 

[[Image:EDA-CBA synthesis.jpg|650x287px|EDA-CBA synthesis.jpg]]

0.055 g 2-amino-1-ethanol

0.26 g CBA

2.5 ml 25% MeOH

60 oC for 2 days.

Then add 0.138 g PEI18 in 2 ml 25% MeOH, 45 oC for 24 hr.

== Plate 3, 2A1E-CBA-PET18 vs PHP-DSP-PEI18 ==

2A1E: 2-amino-1-ethanol 

 

{| cellspacing="1" cellpadding="1" border="1" style="width: 580px; height: 122px;"
|-
| 2A1E-CBA-PEI18, 1:10 
| 2A1E-CBA-PEI18, 1:20 
| 2A1E-CBA-PEI18, 1:30 
| 2A1E-CBA-PEI18, 1:40 
|-
| 2A1E-CBA-PEI18, 1:50 
| 2A1E-CBA-PEI18, 1:75 
| 2A1E-CBA-PEI18, 1:100 
| 2A1E-CBA-PEI18, 1:150 
|-
| PHP-DSP-PEI18, 1:50 
| PHP-DSP-PEI18, 1:75 
| PHP-DSP-PEI12, 1:75 
| PHP-DSP-PEI18, 1:30, 11/10/2011 
|}

36 hours after transfection 

[[Image:2011 11 15 plate 3 s.jpg]] 

 

[[Image:Plate3 11.15.2011.jpg|600px|Plate3 11.15.2011.jpg]] 

[[Image:Plate3 2011 11 15.jpg]]

4 days 

[[Image:2011 11 17 plate3.jpg]] Transfection efficiency is low.

only 1:10 has somewhat good expression

Protocol 2 

PEI-CBA 

== Branched PEI2 (2k). ==

1. Dissolve 20 mg PEI2  in 5 ml MeOH/H2O solution (9:1, v/v) and stirr in a three-necked flask equipped with a condenser at 45âŠC under N2 . 

2. Add  different amount of CBA solution in methanol dropwise into polymer solutions. 

3. Stir the mixture in the dark at 45âŠ C under N2 atmosphere for 24 hr 

3. Add  10% (w/w) excess PEI2 (1 mg/ml) to consume any un-reacted CBA, stir for another 6 h. 

4. Dilute to 10 ml with distilled water and acidified to pH 4 with 10 M HCl. 

5. Dialyse s (MWCO = 3500) against distilled water for 48 h. 

6. Lyophilise for 24 h to give white powder. 

== PEI2âPA ==

N -acylation of PEI2K with PA  Palmitoyl chloride (PA) 

1. Dissolve PEI2 (100 mg) in CHCl3 (100 ml) and stir in a three-necked flask under N2 . 

2. After 30 min of stirrng, add triethylamine (80 ÎŒl) to the solution, stir for 30 min. 

3. Cool the reaction mixture to approx. 4âŠ C. 

4. Add different amounts of PA solution (10, 25 and 50 ÎŒl of 1 mg/ml in CHCl3 ) drop-wise to reaction mix under stirring. 

5. Stir at room temperature for 12 h. 

6. Collect product by precipitation in excess ether. 

7. Lyophilization. 

--[[User:Paul.xu|Paul.xu]] 14:45, 17 November 2011 (EST)

[[Category:Transfection]] [[Category:Paul]]

Synthesis 2A1E-CBA-PEI - Revision history

Paul.xu: moved Synthesis EDA-CBA-PEI to Synthesis 2A1E-CBA-PEI