4 replies to this topic

[leptopelis]

Does anybody have any experience with magnetic-activated cell sorting (MACS)? How efficient and reproducible is it? I would like to use it to isolate adult stem cells from primary cultures because we have no flow cytometer.

leptopelis

Edited by leptopelis, 09 September 2009 - 03:36 AM.

[cotchy]

Im not familiar with MACS but maybe this suggestion is similar,

There is a product called dynabeads by invitrogen and they reckon they are very efficient in separation,

The only catch being they can separate cells only on the basis that a membrane/cell surface protein can be used to separate cells

Basically if your cell type has a known membrane/cell surface protein that all other cells in a mixed population dont have you can purchase antidodies from the same company (they can even make antibodies for you if there are none available for your particular protein),

They then link this to a primary Ab to a secondary antibody which has a metallic element attached to it which can then be separated using a simple magnet held against the side of your eppendof tube or column while flusing out the remaining cell popultion, so no hi-tech equipment is needed.

I've attached the link to their home page

http://www.invitroge...s-and-Uses.html

hope this helps,

i found the company very helpful whern i enquired about the products unfortunately we could not use it due to having no cell specific cell surface protein so try give them an email.

[leptopelis]

Thank you! I have known about Dynabeads but I did not know that there is a conjugation method that allows release of the antibody (and the cells bound to them). I'll contact the company.

[Chinmaya Mahapatra]

https://docs.google.com/document/d/17_zjMinAt8Wob3N5nJwallgys8GugxYkcRj7zFHJWIw/edit

[Natalia KH]

I was considering to use Dynobeads, but than decided to test microbeads instead, since the literature review pointed out that cells experience much less damage with microbeads, which is of course good for the downstream applications. I am using none of them yet, but possibility of cell damage, I think, must be considered.