Frequently Asked Questions
Q: Which kind of instrument is capable of reading PAIA assays?
A: You can either use fluorescence plate readers or automated microscopes for measuring PAIA assays. Both types of instruments provide a very similar performance and are comparable in terms of speed. Plate readers need to have fluorescence bottom reading and shall be able to reliably measure 384-well plates. Please do not hesitate to contact us in case you are not sure about your specific plate reader. Regarding the microscopes we recommend the Cellavista and NyONE imagers from SynenTec GmbH (www.cell-imager.com).
Q: My plate reader can only read fluorescence from the top. Can I still run PAIA assays on it?
A: This is not possible because the instrument will then also detect the fluorescence on the beads and this gives erroneous results.
Q: Which wavelengths are you using in the PAIA assays?
A: We are using the same dyes for all our assays. Optimal excitation is at 640 nm and emission at 665 nm. For microscopes we recommend the filter setting for Cy5 dyes (far red channel in Synentec Imagers).
Q: Is it possible to only use a part of the 384-well plate?
A: Yes. If you have used a part of the plate and want to use the remaining wells later, seal the plate with a foil and keep the plate and the remaining buffer in its original packaging in the fridge at 4° C/38° F. The assay can be stored for at least 4 weeks under these conditions.
Q: Do I need to make a calibration on each plate?
A: If you want to obtain an accurate quantification it is always advisable to do so. You can use calibration data from a different plate, provided that this plate has been processed and measured in the same way. For a qualitative ranking it is also possible to work without any calibration at all.
Q: I would like to incubate several PAIAplates sequentially and then measure them all at the same time. Is this possible?
A: Yes, you can incubate the assays sequentially and then read the plates in one go.
Q: Do you have recommendations for shakers for running PAIA assays?
A: We have quite some experience with orbital shakers, but not with all the shakers on the market. In general mixing speeds of at least 1.400 rpm are necessary for shakers with an orbit of 3 mm and 1.800 rpm for shakers with an orbit of 2 mm. Otherwise you may have to extend the incubation time. Please contact us if you need advice on shakers.
Q: How do I choose the right assays for my application?
A: Please first check which assay is capable of measuring your molecule of interest. Then you should have a look at the concentration range that you expect in your samples. We are offering assays for different concentration ranges, so that you can skip dilutions steps that you often need to perform when using other methods.
Q: Do you offer custom development for special assays?
A: Yes, we offer development services. Development of PAIA assays is less tedious than e.g. setting up ELISA assays and saves internal resources. Please contact us at email@example.com
Q: Are PAIA assays also available in 96-well format?
A: PAIA plates are always 384-well plates to reduce sample volume, reagents and cost per sample. If your workflow is based on 96-well plates you will have to transfer them into our 384-well plates, which can easily be done with either multichannel pipets or with (semi-)automated pipetting robots. You can combine samples from four 96-well plates onto one PAIAplate. We can help you find the easiest way to establish a smooth workflow.