Highly compatible plates for HTRF®
The right plate for a successful assay
Microplates are an important element of FRET signal detection. While most available microplate brands are compatible with HTRF technology and products, a number of parameters are critical to the quality of results.
- FIA white plates from many suppliers remain the most appropriate standard. The signal to noise ratios obtained using white microplates do not differ greatly from those of black plates. Absolute counts, however, are significantly higher in white plates, thereby improving count statistics and subsequent assay reproducibility and sensitivity. NOTE: The use of white plates is mandatory for some assays and highly recommended when detection is performed using certain instruments, in particular on monochromator-based plate readers (Please refer to the package inserts).
- Background levels may differ substantially among various plate types. Tissue-culture treated, or surface-treated plates may also be used.
- Glass bottom plates produce suboptimal results and should not be used.
All the benefits of HTRF with ELISA plate ease of use
Taking these considerations into account, Cisbio’s 96w and 384w low volume microplates have been carefully designed to provide the best signal detection environment possible to all HTRF assays. They can be used in a no-transfer protocol with suspension cells or with a transfer step for adherent cells. These plates can be read on all HTRF certified readers, thus making their implementation in your experiments really easy
Other plates produced excellent results during our validation process for a broad range of HTRF assays. We have listed them in table. 1 below.
|Corning||96-w half area TC treated||100 µL||White||3688|
|Greiner||96-w half area TC treated||100 µL||White||675083|
|Greiner||384-w low volume||15-20 µL||White||784075|
|Greiner||384-w||80-100 µL||White – TC* – sterile||781073|
|Greiner||384-w||80-100 µL||White – TC* – sterile||781080|
|Greiner||384-w low volume||15-20 µL||White – TC* – sterile||784080|
|Greiner||1536-w||5-10 µL||White – TC* – sterile||782080|
|Greiner||1536-w||5-10 µL||White – TC* – sterile||782078|
|Greiner||1536-w||5-10 µL||White, sterile||782073|
|Greiner||1536-w||5-10 µL||White – TC* – sterile||782093|
|NUNC||384-w low volume||15-20 µL||White, NT||264706|
Table 1: a selection of recommended plates and corresponding volume per well
Whatever the plate type, assay volume should match well volume as closely as possible so that focalization of the reader excitation beam gives maximum energy delivery (Fig. 1) . For instance, when assay volumes are equal to or lower than 20 µl, it is better to use 384 low-volume or 1536-well plates rather than regular 384-well plates.
Matching culture containers with signal detection formats
In case your experimental or culture conditions require you cultivate your cells in a specific flask or plate format prior to performing an assay, you can still benefit from an optimal reading by adapting the transfer step between culture format and 96w or 384w reading plate.
This Technical Note will help you performing that transfer step as smoothly as possible.