Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


protocol for liposome preparation

protocol for liposome preparation - Protocols and Methods Forum

protocol for liposome preparation - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 11-29-2008, 06:16 PM
michael nelson
Guest
 
Posts: n/a
Default protocol for liposome preparation



Hey, guys,

I have some difficulties preparing for clear liposomes solutions. Here is what I did, please advise.

*I start off with 0.1g/ml phosphatidylcholine chloroform solution. I thenmove it into a round bottomed flask and dry it in a rotary evaporator. However, instead of getting thin films on the surface, what I end up with is alarge chunk of light yellowish solid. Does that mean I have to use a larger volume of organic solvent?

I then dissolve them into buffer and sonicate. Finally, I got some milky suspension. I assume they are multimellar vesicles. My advisor told me that Icould sonicate to make them unilamellar and make the solution goes clear. I tried, but didn't work.

Could anyone here tell me how to prepare the unilamellar vesicles in a clear solution? More details are better.

Thanks!






Reply With Quote
  #2  
Old 11-29-2008, 08:22 PM
WS
Guest
 
Posts: n/a
Default protocol for liposome preparation

Hi Michael,

you might try this: dissolve PC in EtOH and inject into 20 to 50 fold
volume of buffer with an insulin syringe (or the narrowest needle you
can get) while vortexing. If the EtOH might cause problems downstream,
dialyze against buffer.

Worked well for PC/Ceramide vesicles for me.

best regards,

Wo
Reply With Quote
  #3  
Old 12-01-2008, 12:34 PM
Dr Engelbert Buxbaum
Guest
 
Posts: n/a
Default protocol for liposome preparation

Am 29.11.2008, 14:16 Uhr, schrieb michael nelson <[Only registered users see links. ]>:



There is the classical method of pressing these multilamellar vesicles
through a narrow-pore membrane. However, more consistent results you will
obtain by dissolving the lipids with detergents as mixed micelles and then
striping the detergent away with polystyrene beads (e.g. Biobeads SM2).
Rigaud's group has done extensive work on this, check their papers.
Reply With Quote
  #4  
Old 12-03-2008, 02:18 PM
jh
Guest
 
Posts: n/a
Default protocol for liposome preparation

Excuse me, may you explain more detail for "PC".
Whether the liposome is prepared for cell transfection experiment?

Jianhao

2008/11/30 WS <[Only registered users see links. ]>:

Reply With Quote
  #5  
Old 12-03-2008, 09:00 PM
WS
Guest
 
Posts: n/a
Default protocol for liposome preparation

On 3 Dez., 15:18, jh <[Only registered users see links. ]> wrote:
Hi,

PC means phosphatidylcholine. I haven't used them for transfection but
to transfer a fluorescently labeled ceramid into fibroblasts.

Regards,

Wo



Reply With Quote
  #6  
Old 12-05-2008, 02:26 AM
jh
Guest
 
Posts: n/a
Default protocol for liposome preparation

Thanks.
I wonder whether anyone prepare liposome by self for cell transfection
experiment?Cause ,I thought, this may help for someone who do not have
much fund.

rgds!

jianhao

2008/12/4, WS <[Only registered users see links. ]>:

Reply With Quote
  #7  
Old 12-05-2008, 04:53 AM
DK
Guest
 
Posts: n/a
Default protocol for liposome preparation

In article <[Only registered users see links. ].net> , jh <[Only registered users see links. ]> wrote:

People who don't have funds to afford various lipofection reagents
on the market would probably do well by carefully optimizing
polyethyleneimine-mediated transfection. Lots of reports that suggest
that PEI gives transfection efficiencies comparable to many cationic
lipids.

DK
Reply With Quote
  #8  
Old 12-06-2008, 10:10 AM
jh
Guest
 
Posts: n/a
Default protocol for liposome preparation

thanks
Do you have the protocol in detail?
Or Would you like to recommendate one?

rgds!

Jh

2008/12/5, DK <[Only registered users see links. ]>:

Reply With Quote
  #9  
Old 03-18-2011, 10:28 AM
Pipette Filler
Points: 52, Level: 1 Points: 52, Level: 1 Points: 52, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2011
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: protocol for liposome preparation

Hello,
does anybody have any idea for PC clarification? i have it in 60% concentration. I already washed it with acetone. but i am searching for any other possibilities.

Thank you.

CK
Reply With Quote
Reply

Tags
liposome , preparation , protocol


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
A protocol for GST preparation and protein purification Arabidop Molecular Biology Articles and Protocols 2 04-24-2010 04:50 AM
protocol for preparation of actomyosin heartthrob Protocols and Methods Forum 0 09-14-2005 09:05 AM
protocol for preparation of actomyosin heartthrob Protocols and Methods Forum 0 09-14-2005 09:01 AM
protocol for preparation of actomyosin heartthrob Protocols and Methods Forum 0 09-14-2005 09:01 AM
Sample Preparation Workshop - Ottawa - November 16, 1st Announcement. Victor Benham Forum Chemie 0 11-05-2004 01:56 AM


All times are GMT. The time now is 02:14 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.15393 seconds with 16 queries