FLIPR Calcium 3 Assay Kit

Superior Detection for Diversified Targets

• No wash steps
• Adherent and non-adherent cell types
• Higher signal intensity
• Improved data quality
• Compatible with more targets

Introduction

The FLIPR Calcium 3 Assay Kit provides a universal method for detecting changes in intracellular calcium concentration in a simple and reliable homogeneous assay format. Specifically, this fast and simple fluorescence-based assay yields maximum signal intensity, thereby allowing the researcher to accurately and easily detect intercellular calcium fluxes across a broad spectrum of targets. Further, the FLIPR Calcium 3 Assay Kit is designed to produce superior results from chemokine and small peptide targets, as well as transient, endogenous, orphan and low-expression receptors that have been difficult to assay with standard assay formats.

FLIPR assays for GPCRS

Calcium mobilization assays for screening GPCR targets are preferred techniques in drug discovery because they generate high quality leads. The Molecular Devices FLIPR system is the method of choice among pharmaceutical and biotechnology companies for screening calcium flux assays in high throughput. The Molecular Devices FlexStation™ is also commonly used in assay development, target validation and lower throughput applications.

Optimized assays

To simplify assay development and optimize performance on both FLIPR and FlexStation, Molecular Devices has developed a series of no-wash assay kits. The combination of proprietary chemistry and a no-wash protocol causes less cellular disruption than traditional methods to generate consistent results and strong signals. FLIPR Assay Kits provide the easiest way to get the best performance from your FLIPR or FlexStation system.

Patented technology

FLIPR Assay Kits from Molecular Devices employ a quenching dye to reduce background fluorescence and improve the signal-to-noise ratio. The patented quench technology (U.S. patent number 6,420,183, EPO patent number 0 906 572) is offered to drug discovery and life science researchers exclusively by Molecular Devices, through the purchase of FLIPR Assay Kits.

Universal format

Using the patented quench technology and advances in reagent chemistry, the FLIPR Calcium 3 Assay Kit provides an improvement in signal strength and data quality over previous methods. The assay protocol results in fewer spontaneous calcium fluxes and unresponsive cells. When compared with the standard FLUO-3AM dye incubation-and-wash procedure, assay signal intensity using the FLIPR Calcium 3 Assay Kit is substantially higher and more distinct, resulting in superior detection. Compared to the standard FLUO-4 AM dye incubation-and-wash procedure, the kit produces equivalent or greater signal intensity (Figure 1) and better well-to-well uniformity, resulting in superior data quality and a higher Z´-factor. This allows the researcher to accurately and easily detect intracellular calcium fluxes even from chemokine and small peptide targets, as well as transient, endogenous and low-expression receptors that have been difficult to assay with standard methods. Strong signals are attained in cases that may previously have yielded weak signal intensity or no peak at all. Thus, the FLIPR Calcium 3 Assay Kit negates the need for having multiple calcium flux assay chemistries for different receptor targets.

		Figure 1. Comparison of signal intensity obtained with FLIPR Calcium 3 Assay Kit vs. FLUO-4 and FLUO-3 wash protocols. Response of CHO cells transfected with M1-muscarinic receptors to 100 nM carbachol as measured on a FLIPR system using the FLIPR Calcium 3 Assay Kit.

Rapid assay development

The streamlined, homogeneous format makes the FLIPR Calcium 3 Assay Kit less labor intensive to run. With the mix-and-read procedure, the cells can be incubated with the reagents and are stable for up to several hours. Rapid analysis of the cells can be followed with detection on a FLIPR system or FlexStation® microplate reader.

Increased throughput

The FLIPR Calcium 3 Assay Kit increases throughput by eliminating cumbersome wash steps. Less preparation time is required for wash buffers and wash calibrations. This successfully eliminates causes for variability in the data, resulting in fewer false positives and negatives. The kit is designed to work at room temperature, which is conducive to automation using stackers or robots. Larger unit volume packaging in the Express kit format minimizes reagent bottle and liquid handling.

Reduced assay failures

Cell detachment, lowered cell competency and incomplete dye washing are common problems that are easily avoided by using the FLIPR Calcium 3 Assay Kit. In addition, the elimination of potential dispensing and washing errors, or associated equipment failures, further ensures the integrity of screening operations.

Ordering Information

To place an order, call your local Molecular Devices Office or Molecular Devices Distributor.