As starting dilution in HS ELISA, with all three items, we recommend 1/5.000 (200 ng/ml).
This dilution will be close to saturating dilution in many HS ELISA systems that with good
antibodies will yield close to maximum sensitivity. In some rapid ELISAs more
concentrated conjugates (up to 1/2.000 but unlikely more concentrated than 500 ng/ml) can
be applied within the same 30-min. or shortened 20-15-min. incubation time.
In practice, however, developmental situation with SA-PolyHRP does frequently look
Many researchers, driven by "pressure" of high backgrounds, tend to dilute SA-PolyHRP
very strongly applying conjugates at working strength dilutions down to 1:10.000 - 1:20.000
They of course lose some sensitivity but at the same time obtain more "comfortable"
situation with backgrounds and gain advantage of having on the total very economical
reagent consumption levels.
This however creates another difficulty when working with stock PolyHRP conjugate
solution. Making daily working dilutions necessitates dispensing minor microliter volumes.
Practically, working strength conjugate dilution in such cases must often be produced
through executing intermediate dilution(s).
This makes daily developmental work with PolyHRP stock solution either less precise and
less reproducible or less economical, or quite precise but more cumbersome and, again, less
Suggestion to use SA-PolyHRP Stabilizers addresses this experimental circumstance.
Stabilizers give the possibility of creating intermediate conjugate dilutions that will be long
time stable at regular storage
(+2°C/+8°C) and handling/working (+18°C/+25°C) temperature regiments. Preparing stable
intermediate formulation in the range e.g. 500X - 10X of final working strength - for
further making daily working dilutions respectively 1/500 - 1/10 - will make developmental
work with PolyHRP much more convenient, more precise and more economic.
That is why we currently offer PolyHRP also in pre-diluted stable at
+4°C/+25°C format. Three different versions (50, 10 & 2 µg/ml) of
SA-PolyHRP are available within convenient choice of flexible packaging in diverse sizes.
This option will increase level of precision and overall reproducibility in developmental
work with PolyHRP. Working with stable pre-diluted PolyHRP conjugates is strongly
recommended when running at least primary development.