Further, it has also been discovered that aequorin assays like PerkinElmer’s AequoScreen^® can be read on fluorescent-based readers such as the FLIPR^TETRA®.

A recent poster demonstrated that “signal intensity for…selected catalog aequorin cell lines is strong enough to allow its measurement by the non-luminescence FLIPR^TETRA®” reader. The research team found “several examples of aequorin signal detection with the FLIPR^TETRA®, generating high-quality data, with Z’ values compatible for HTS use of AequoScreen assay on the FLIPR^TETRA®.”

The reagents used in the study included “double transfected aequorin cell lines, stably expressing both mitochondria-targeted aequorin and a GPCR,” as well as one of the “AequoScreen parental cell lines,” which can be used by customers to introduce their own receptors:

• Histamine H₁ AequoScreen cell line
  (PerkinElmer #ES-390-A)
• Vasopressin V_1B AequoScreen cell line
  (PerkinElmer #ES-362-A)
• Vasoactive Intestinal Peptide VPAC₁ AequoScreen cell line (PerkinElmer #ES-273-A)
• CHO-K1 (+Gα₁₆) AequoScreen Parental cell line
  (PerkinElmer #ES-000-A24)

Luminescence cell lines intense enough for FLIPR^TETRA®

PerkinElmer’s AequoScreen (aequorin) and PhotoScreen™ (Photina^®) portfolios are a good match for FLIPR^TETRA®. Simply turn off the excitation light of the FLIPR^TETRA® and easily generate high-quality data with exceptional Z’ values using these validated cell lines. In fact, tests showed equivalent luminescence on FLIPR^TETRA® and LumiLux^® for select cell lines (Figure 1).

A further advantage of utilizing these cell lines is that there are no wash steps following substrate loading, enabling faster results as well as the potential for easier automation for high throughput screening (HTS) workflows. It is important to note that reading in luminescence and fluorescence modes has also exhibited equivalent EC₅₀ values; however, there may be differences for some receptors.

• Receptor Ligand Binding
• Receptor Membranes
• GPCR Cell Lines
• cAMP Quantitation
• Phosphorylation of ERK
• GTP Binding
• Reporter Gene Assays
• Intracellular Calcium

• Reagents and Assays
• Photoproteins
  
• Cell Lines
• Radioligands
• Detection Instruments
• Automated Liquid Handling
• High Content Screening
• Microplates
• Assay Development & Labeling

• MicroBeta²™ LumiJET™
• EnSpire™ Alpha Plate Reader

Figure 1. Pharmacology. Luminescence Assay FLIPR^TETRA® vs. LumiLux

384-well Histamine H₁ (above) and Vasoactive Intestinal Peptide (VPAC₁) (below) receptor AequoScreen receptor cells were analyzed on the FLIPR^TETRA® and the LumiLux in 384-well format using a flash luminescence aequorin assay. EC₅₀ values obtained on both readers and in both assay modes were equivalent. In adherent mode, frozen cells were thawed and plated before loading with coelenterazine prior to analysis.

Luminescence cell lines intense enough for FLIPR^TETRA® (with the aequorin option)

In addition to the possibility of using the FLIPR^TETRA®, a majority of PerkinElmer’s AequoScreen and PhotoScreen cell lines could be suitable for reading with the new FLIPR^TETRA® (with aequorin option) as this has improved sensitivity dedicated to both luminescence and fluorescence readings.

Make your own aequorin cell lines

FLIPR^TETRA®-compatible luminescence cell lines expressing GPCRs of interest can also be generated using parental AequoScreen cell lines or by transfecting aequorin plasmids into a desired recombinant GPCR-expressing cell line. The choice is yours depending on your assay requirements.

PDL-coated microplates can improve your performance

Ideal for fluorescent and/or luminescent assays in any FLIPR^® model, PerkinElmer ViewPlates^®, tissue culture (TC) treated and poly-D-lysine (PDL)-coated plates are available in 96-, 384- and 1536-well formats. Researchers can have confidence in their results with these high-quality microplates.

Want more information?

• Click here to downlaod a copy of the poster.
• Click here to download information about aequorin on FLIPR^®.

©2008-2009 PerkinElmer, Inc. All rights reserved. The PerkinElmer logo and design and AequoScreen, LumiLux and ViewPlate are registered trademarks, and EnSpire, LumiJET, MicroBeta² and PhotoScreen are trademarks, of PerkinElmer, Inc. or its subsidiaries, in the United States or other countries. FLIPR and FLIPR^TETRA are registered trademarks of MDS Analytical Technologies. Photina is a registered trademark of Axxam, Inc. All other trademarks not owned by PerkinElmer, Inc. or its subsidiaries that may be depicted herin are the property of their respective owners.