HTRF® microplate recommendations
Photo courtesy of Greiner
Most available microplate brands are compatible with HTRF technology and products. Nevertheless, background levels may differ substantially among various plate types. FIA black plates from many suppliers remain the most appropriate standard. Tissue-culture treated, or surface-treated plates may also be used. Glass bottom plates produce suboptimal results and should not be used. The following plates produced excellent results during our validation process for a broad range of HTRF assays:
| Provider | Format | Recommended volume | Type | Cat# |
|---|---|---|---|---|
| Greiner | 384-w | 80-100 µL | Black | 781076 |
| Greiner | 384-w | 80-100 µL | White | 781075 |
| Greiner | 384-w low-volume | 15-20 µL | Black | 784076 |
| Greiner | 384-w low-volume | 15-20 µL | White | 784075 |
| Greiner | 96-w | 160-200 µL | White - TC* - sterile | 655083 |
| Greiner | 96-w | 160-200 µL | Black | 655209 |
| Greiner | 96-w | 160-200 µL | Black - TC* - sterile | 655086 |
| Greiner | 96-w | 160-200 µL | Black - TC* - sterile | 655079 |
| Greiner | 96-w | 160-200 µL | White - TC* - sterile | 655073 |
| Greiner | 384-w | 80-100 µL | Black - TC* - sterile | 781086 |
| Greiner | 384-w | 80-100 µL | Black - TC* - sterile | 781079 |
| Greiner | 384-w | 80-100 µL | White - TC* - sterile | 781073 |
| Greiner | 384-w | 80-100 µL | Black | 781209 |
| 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 | 783075 |
| Greiner | 1536-w | 5-10 µL | Black | 783076 |
| Greiner | 1536-w | 5-10 µL | White - TC* - sterile | 782080 |
| Greiner | 1536-w | 5-10 µL | Black - TC* - sterile | 782086 |
| Greiner | 1536-w | 5-10 µL | White - TC* - sterile | 782078 |
| Greiner | 1536-w | 5-10 µL | Black, sterile | 782092 |
| Greiner | 1536-w | 5-10 µL | White, sterile | 782073 |
| Greiner | 1536-w | 5-10 µL | White - TC* - sterile | 782093 |
| Corning | 96-w | 160-200 µL | Black, NT | 3915 |
| Corning | 96-w | 160-200 µL | White, NT | 3912 |
| Corning | 96-w half-area | 80-100 µL | Black, NT | 3694 |
| Corning | 96-w half-area | 80-100 µL | White, NT | 3693 |
| Corning | 384-w | 80-100 µL | Black, NT | 3573 |
| Molecular Devices | 96-w half area and half-volume | 30-40 µL | Black, NT | 42-000-0117 |
| NUNC | 384-w low-volume | 15-20 µL | Black, NT | 264705 |
| NUNC | 384-w low-volume | 15-20 µL | White, NT | 264706 |
| Perkin Elmer | 384-w low-volume | 15-20 µL | White proxiplates | 6008289 |
| Perkin Elmer | 384-w low-volume | 15-20 µL | White proxiplates | 6008280 |
| Perkin Elmer | 384-w low-volume | 15-20 µL | White proxiplates | 6008281 |
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. 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.
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, such as IP-One, and highly recommended when detection is performed using certain instruments, in particular on monochromator-based plate readers.