How to dilute an antibody? - (May/22/2008 )

Hello all,

I need to know the best way to dilute an antibody, as i have having a lot of inconsistent results with my assays.

I only need very small amounts as i am using it for an ELISA. My working dilition is 1/120,000 of my original concentrate tube. I do not know the buffer the original is in, as it is not available commercially.

So i clearly need to dilute the original tube down first, as i only need a couple of ml maximum at any one time.

So what is the best way to dilute down? a 1/100? 1/1000? and, is picking up 1ul accurate? so therefore should it be 5/500 and so on? And as i dont know the buffer, what is the best buffer to dilute in? And should it be able to be stored as long as if it had not been diluted?

I am sure my assays are inconsistent because od this factor, so any help will be GREATLY appreciated!
Thank you...

-Flour Power-

QUOTE (Flour Power @ May 22 2008, 11:45 AM)

The easiest way is to take out an easily measurable volume and it also depends of your pipets

Bets would be a 1/500 dilution so 2microliters in 1ml and do your other dilutions from there.

You might also keep the diluted antibody (first dilution) in your fridge and always prepare a fresh dilution from this to minimize the dilutions factors

-Jipes-

QUOTE (Jipes @ May 22 2008, 04:54 AM)

I need to know the best way to dilute an antibody, as i have having a lot of inconsistent results with my assays...

Check some of these results..

http://search.vadlo.com/b/q?sn=158621799&a...ution&rel=0

..

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