I am searching for analytical standards for beta-estradiol and testosterone to use in spike-recovery experiments to validate ELISA kits. What form of standard is best to use? I see that most are soluble in ethanol or acetonitrile and I am concerned about this solvent interfering with the assays. Preferably I would like to dissolve in water or the diluent. Thanks.
I saw some "water soluble" steroid packed in cyclodextrin. Like Sigma H0396 or E4389.
Thank you Attila, I'm not sure if these would be suitable for use as analytical standards but I will look into it further. Cheers, Mazen
Hi Angus,
Thanks for the info, very helpful!
I have spoken with our Endocrine lab and they also do the same, they dissolve the steroid in ethanol and then dilute this well with the assay buffer.
All the best with your research, very interesting.
Kind regards,
Mazen
Hi Mazen,
up to 1% organic solvents (DMF or EtOH) in your standards should be OK, so a 100:1 dilution of the ethanolic steroids in diluent should work.
Kind regards, Jürgen
Thanks very much Jürgen. I will keep that in mind when designing my experiments.
Kind regards,
Mazen
... and at the end, you have to make sure, that you have equal conditions for samples and standards (and QCs as well). By using monoclonals, they can be affectet differently by solvents and other assay conditions.
You should run a "disturbance protocol" for your assay. Checkerboard titrtration: antigen from row A - H (use your current concentrations and blanks), disturbing factor(s) from column 1 - 12 (use the expected final concentration range either in a mixture of ethanol or acetonitrile or alone)
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