I'm trying to measure IgG level from plasma samples that I collected weekly from my animal experiment to see how IgG level changes throughout the study.
Since the level will be different in samples from earlier time point and later time point,
I'm wondering if I can use two different dilution factors have all of the samples fall under standard range.
If I set one dilution factor based on high level samples, rest of time points will be below standard range.
What would be best way to set up this assay?
Hi,
My suggestion for you to perform the checkerboard titration for antibody. After, you can start with 1:250 dilution and can go up to 1:192000. Now to fix the concentration of antibody and antigen take 20% more than the plateau value.
Ps: ELISA is sensitive to detergents and it can have a great impact on the results. Usual detergents used are Triton X-100 or more usual Tween 20 as low as 0.05%.
which type or subclass of IgG you try to measure?
As you know
The IgG class of antibodies is composed of four different subtypes of IgG molecules called the IgG subclasses. These are designated IgG1, IgG2, IgG3 and IgG4.
ELISA is sensitive and it can have a great impact on results.
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