Q about checkerboard titrations

[Elle88]

Hello, I'm Elle and i'm new to this site!

 

I'm currently doing some research on respiratory diseases.

 

I was wondering would anyone be able to give me some advice on what a "checkerboard titration is exactly" and how I would go about doing one with relation to a conjugate antibody?! I've been looking everywhere and don't seem to be finding much!

 

From what I have found, I think a checkboard titration is used to find the optium concentration of the reagent, but i'm still not entirely sure what all it involves from a practical point of view!

 

Elle

[CharonY]

From what I have found, I think a checkboard titration is used to find the optium concentration of the reagent,

This is basically correct. It is used to find an optimum concentration or mixture of reagents. It is normally used in ELISA assays (I assume that was your starting point) but in variations it can be used in all concentration based assays on plates.

In the simplest case you can imagine one reagent(e.g. primary antibody) diluted 1:1 1:5 1:10 and so on across the plate and a dilution series with the second reagent (e.g. secondary antibody) down the plate.

So in the top left (A1) you would have a mixture of 1:1 primary to 1:1 secondary antibody, in the well left to it (B1)1:5 primary to 1:1 secondary, and in the well below the top left (A2) 1:1 primary to 1:5 secondary.

If you fill the plate you then can easily see which ratios are in the optimum for your experiment, or you can determine the possible range of your measurement (by using a dilution series of standards) and so on.

[Elle88]

Thank you for your help, it was very useful!

 

Elle