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How to remove azide from antibody


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#1 TracyDuke

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Posted 17 August 2009 - 01:22 PM

Does anybody know how to remove sodium azide from the comercial antiboy solutions? I know there are two ways: dialyzing or exchanging over a column. Could anyone provide more detailed information? Thank you all!

#2 cellcounter

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Posted 17 August 2009 - 02:04 PM

Does anybody know how to remove sodium azide from the comercial antiboy solutions? I know there are two ways: dialyzing or exchanging over a column. Could anyone provide more detailed information? Thank you all!

I would go for a Dialysis using tubing/cassette that has something like 10KD cut-off. This will let Azide pass through while retaining antibody and BSA in your sample.

#3 miBunny

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Posted 17 August 2009 - 05:04 PM

IF you are working with a volume under 500 uL, the microcon filter is the easiest. Hopefully, I've successfully loaded the micron filter bulletin which walks through the entire procedure. If not, heres a link.

http://www.millipore...des/tech1/99394

If you have more than 500 uL, you probably will need to do a dialysis. You will need dialysis tubing with a molecular weight cut-off of around 12,000-14,000. You regenerate the tubing/membrane and clip off one end. Add the material to the tube and seal the other end. Place the material in an excess of PBS and let it sit (with stirring) at 4 degrees. Change the PBS a few times and give it at least an overnight incubation (24 hours is better). Here are a few links with more information

http://www.sigmaaldr...dium-azide.html
http://www.rockland-...oval_Method.jsp

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