5 replies to this topic

[AllenChiu]

hey guys，what do you think the Anti-Flag M2 agarose beads is made of?
If it is made of regular agarose,why wouldn't it melt when I boiled the beads with loading buffer?
Well,actually I want to make some homemade agarose beads,which haven't been crosslinked to any protein A/G or antibody.
Because I suspected the non-specific binding of IP is caused mainly by the agarose part of the beads,instead of the protein A/G or antibody.
So I want to preclear the lysis with large amount of cheap homemade agarose beads?
Do you think it make any sense?What do you think?
Thanks

[mdfenko]

the way to make acrylamide beads for chromatography is to polymerize acrylamide at the desired concentration and degree of crosslinking then homogenize it in a large volume of water or buffer and let it settle.

agarose should be about the same.

[phage434]

For agarose, the usual approach is to put droplets of hot agarose into cool mineral oil with a pipet tip.  You can control the size by choice of tip and how up detach the droplet from the pipet.

[AllenChiu]

mdfenko, on Mar 18 2010, 04:00 AM, said:

the way to make acrylamide beads for chromatography is to polymerize acrylamide at the desired concentration and degree of crosslinking then homogenize it in a large volume of water or buffer and let it settle.

agarose should be about the same.

Thanks，It’s very kind of you

[AllenChiu]

phage434, on Mar 18 2010, 08:54 AM, said:

For agarose, the usual approach is to put droplets of hot agarose into cool mineral oil with a pipet tip.  You can control the size by choice of tip and how up detach the droplet from the pipet.

Your solution is very useful,thanks!

[lovebugs1]

hi allen,

Did you successfully make your own agarose beads? I would like to make my own beads too.
I was wondering if you have a protocol to send to me. Thank you.