Screening For Lectins in bacteria - Help !!! I need help from anybody who had experienced (Aug/24/2009 )

Dear All,

I am doing a project in screening for lectins in Bacteria. I am not a biochemistry or immunology background.
From literature, the screening protocols needs column coupled with sugars. I had done the sugar coupling but how do I test the efficiency of my column?

Also, I had a trouble in cell breaking. Since I suspect the lectin is either membrane bound or reside inter cellular, what would be a most efficient way to break the cell wall and release the lectins within? My bacteria is a gram -ve BSL-2 organism. I found out that literature did mentioned about ultrasonic vibrations. The problem is should I use a "water bath type" sonicator or "probe" type sonicator?

Sorry for my noobness but I really got stuck and got no idea on how to deal with this.

Appreciate your help and comments.

Thanks

Adrian.

check out the handbook found on this webpage:

ge healthcare affinity chromatography handbook


look at the bound lectins and apply it to your problem.

also, i'm pretty sure they meant a probe sonicator.

Dear mdfenko,
Thank you for your reply.

The book's protocol is different from my reference literature. In the book there is mention do not magnetic stirring the agarose but however the GE handbook did mention there not to stir it when I try to activate the gel for ligands coupling. Any idea?

Thank you.

you can put it in a tube and rock it or use an end-over-end rotator to maintain the suspension.

Thank you for your suggestions, dear mdfenko.



I guess I had wasted some portion of my gel due to ignorance...


Now still searching for an economic sonicator for cell breaking...we are lacking of money now.

adrian kohsf on Sep 3 2009, 04:15 AM said:

Hi Adrian,

we at Hielscher offer a lot of equipment at favorable lease conditions with minimum lease terms starting at a month. So you could do your research for a month or two and return the device - no questions asked . Of course all lease payments are credited towards purchase.

If this is of interest to you, pls have a look at our ultrasonic laboratory devices.

Have a nice day!