Paul.xu at 01:30, 1 August 2014

2014-08-01T01:30:12Z

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== Synthesis of PHP Agmatine ==

50 ul PHP in DMSO + 10 ul 0.5 ug/ul agmatine in water 

AG not soluble in DMSO, form ppt 

+ 1 ml isopropanol --> spin, reomve liquid 

+ 100 ul H2O to dissolve 

+ 5ul 100 mg/ml DTBP-->see turbid, slowing become transparent 

+ 5ul 100 mg/ml DSP--> turbid, let react at 25 oC o/n 

{| width="503" cellspacing="1" cellpadding="1" border="1"
|-
| 
| Sample desc 
| Note 
|-
| 1 
| 7/19/14 PHP-DSP-PEI18, purified, 1:10 
| 
|-
| 2 
| 7/19/14 PHP-DSP-PEI18, purified, 1:30 
| 
|-
| 3 
| 7/19/14 PHP-DSP-PEI18, purified, 1:50 
| 
|-
| 4 
| 7/19/14 PHP-DSP-PEI18, unpurified, 1:10 
| 
|-
| 5 
| 7/19/14 PHP-DSP-PEI18, unpurified, 1:30 
| 
|-
| 6 
| 7/19/14 PHP-DSP-PEI18, unpurified, 1:50 
| 
|-
| 7 
| PHP-DSP-Agmatine, synthesis in water 
| 
|-
| 8 
| PHP-DTBP-Agmatine, synthesis in water 
| 
|-
| 9 
| PHP-DSP-Agmatine, synthesis in DMSO, white ppt 
| 
|-
| 10 
| PHP-DSP-Agmatine, synthesis in DMSO, oil after water extraction 
| 
|-
| 11 
| PHP-DSP-PEI18 from 6/15/14, 1:10 
| 
|-
| 12 
| PHP-DSP-PEI18 from 6/15/14, 1:10 
| 
|-
| 13 
| L2K 
| 
|}

Picture  2 Days post transfection 

[[Image:PHP AG 7 23 14.JPG]] 

Picture 5 Days post transfection 

[[Image:Chitosan PHP AG 7 25 14 1.JPG]]

Picture 2 Days post transfection 

[[Image:PHP-PEI18 vs L2K 7 22 14.JPG.JPG]]

Picture 4  Days post transfection 

[[Image:PHP-PEI18 vs L2K 7 23 14.JPG]] 

Picture 5 Days post transfection 

[[Image:Chitosan PHP AG 7 25 14 2.JPG]] 

 

== Conjugate Chitosan to PHP/PEI ==

{| width="756" cellspacing="1" cellpadding="1" border="1"
|-
| Plate Pos 
| Sample # 
| CS sol 
| Conjugate 
| Linker 
| Observation 
|-
| 2 
| 1 
| 20 ul 
| 2 ul 0.4 mg/ml PEI18 
| 
| rowspan="2" | need to add 1 ul 8N HOAc to adjust to pH<7 to dissolve CS 
|-
| 3 
| 5 
| 20     
| 2 ul 0.4 mg/ml PEI18 
| 5 ul 100 mg/ml DTBP 
|-
| 4 
| 2 
| 20
| 2 ul 0.5 mg/ml agmatine 
| 
| rowspan="2" |  form insoluble ppt 
|-
| 5 
| 6 
| 20
| 2 ul  0.5 mg/ml agmatine 
| 5 ul 100 mg/ml DTBP 
|-
| 6 
| 3 
| 20
| 2 ul PHP in DMSO 
| 
| 
|-
| 7 
| 7 
| 20
| 2 ul PHP in DMSO 
| 5 ul 100 mg/ml DTBP 
| 
|-
| 8 
| 4 
| 20
| 2 ul PHP in DMSO + 2 ul 0.4 mg/ml PEI18 
| 
| rowspan="2" | need to add 1 ul 8N HOAc to adjust to pH<7 to dissolve CS
|-
| 9 
| 8 
| 20  
| 2 ul PHP in DMSO + 2 ul 0.4 mg/ml PEI18
| 5 ul 100 mg/ml DTBP 
|-
| 10 
| 9 
| 20 
| 2 ul 0.5 mg/ml agmatine 
| 2 ul 0.5 mg/ml agmatine 
| 
|-
| 1 
| C 
| 
| 
| 
| 
|-
| 11 
| PHP + AG + DSP 
| 
| 
| 
| 
|-
| 12 
| PHP + AG + DTBP 
| 
| 
| 
| 
|-
| 13 
| PHP + AG, ppt 
| 
| 
| 
| 
|-
| 14 
| PHP + AG, oil 
| 
| 
| 
| 
|-
| 15 
| 7/19/14, PHP+DSP-PEI18, purified 1:10 
| 
| 
| 
| 
|-
| 16 
| 7/19/14, PHP+DSP-PEI18, purified 1:30 
| 
| 
| 
| 
|-
| 17 
| 7/19/14, PHP+DSP-PEI18, unpurified 1:10 
| 
| 
| 
| 
|-
| 18 
| 7/19/14, PHP+DSP-PEI18, unpurified 1:30 
| 
| 
| 
| 
|-
| 19 
| 7/20/14, PHP+DSP-PEI18, 1:10 
| 
| 
| 
| 
|-
| 20 
| 7/20/14, PHP+DSP-PEI18, 1:30 
| 
| 
| 
| 
|-
| 21 
| L2K 
| 
| 
| 
| 
|}

7/23/14: Transfection: dilute 1 ul to 20 ul. use 3 ul to transfect 293T cells 

pticture of 7/25/14

[[Image:Chitosan PHP AG 7 25 14.JPG]] 

7/26/14 

[[Image:Chitosan PHP AG 7 26 14.JPG]] 

7/27/14 

[[Image:Chitosan PHP AG 7 27 14.JPG]]

7/29/14

[[Image:Chitosan PHP AG 7 29 14.JPG]]

7/31/14

[[Image:Chitosan_PHP_AG_7_31_14.JPG]]

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

Synthesis PHP-Agmatine - Revision history

Paul.xu at 01:30, 1 August 2014