How to separate A/G beads from the attached antibodies?

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viralscience
viralscience's picture
How to separate A/G beads from the attached antibodies?

I want to pull down my vesicle of interest with antibody coated A/G beads, but, the problem is I want to separate the vesicle from the beads and use that vesicle for other purpose. Does anyone know if there is way to get stuff off the beads without heating? Also, I want to avoid using magnetic beads.

Thanks

VS

Chin Fen Teo
Chin Fen Teo's picture
Hi viralscience,

Hi viralscience,

One way to do it is to incubate with 0.1 M glycine (pH2.5 to 3) at ambient temperature for 5 to 10 min. However, neutralization of your sample (typically, with 1/10 volume of 1 M Tris, pH 8) immediately after acidic elution is needed.

Good luck.

viralscience
viralscience's picture
 Many Thanks!

 Many Thanks!
 Btw, do you have a reference where I can refer to for other details?

VS 

g a
g a's picture
 Hi Viralscience

 Hi Viralscience

You can also use the high salt concentration 1M at acidic pH 2-3.

All the best