4 replies to this topic

[WolfeMD]

Can anyone recommend a good extraction buffer that is compatible with ELISA that will work for Arabidopsis thaliana?

I have previously used SDS based buffers to do Western Blots, but SDS isn't compatible with ELISA because it denatures antibodies. I have had extremely limited success trying to use PBST (phosphate buffered saline with Tween-20).

Any advice would be helpful.

Cheers,

Marnin

[sgt4boston]

After you do the extraction could you not change buffers by dialysis?

[WolfeMD]

sgt4boston, on Apr 9 2009, 03:45 PM, said:

After you do the extraction could you not change buffers by dialysis?

I could potentially do that. However, that is an entire extra step. I would only do that if it is the only way. Additionally, extra steps like that make ELISA less quantitative because there are extra steps with extra opportunity for human error.

Does ANYONE work on plants here?

Help!?

[sgt4boston]

Could you extract; concentrate your material of interest using a Milllipore product (low binding) then dilute your sample? Depending upon the degree of concentration the % of surfactant should be fairly low and not interfere in the test.

[scary]

Hi,

I don't work with arabidopsis but I work on some crop plants like Groundnut, Cotton, Pigeon pea, Sunflower where I use PBST or TBST @ pH-7.6 or  50mM Tris at pH-8.0 as ELISA extraction buffers. Check with these buffers it may help you.

All the best

Scary