What is FLIPR Assay?
FLIPR (Fluorescent Imaging Plate Reader) was developed to perform quantitative optical screening for cell-based kinetic assays. FLIPR incorporates an integrated design, including low-level optical detection, precise temperature control, and precise fluid handling, all in one package.
What is a calcium assay?
Calcium Assay Kit (Colorimetric) ab102505 provides a simple assay to determine calcium concentration within the physiological range of 0.4 – 100 mg/dL (0.1 – 25 mM). In the calcium assay protocol, a chromogenic complex is formed between calcium ions and 0-cresolphthalein. The complex is measured at OD = 575 nm.
How do you measure intracellular calcium levels?
Changes in intracellular calcium concentration can be measured using the calcium sensitive fluorescent ratiometric dye fura-2 AM. This method is a high throughput way to measure agonist mediated calcium responses.
What does FLIPR stand for?
FLIPR
| Acronym | Definition |
|---|---|
| FLIPR | Fluorometric Imaging Plate Reader |
| FLIPR | Fluorescent Imaging Plate Reader |
| FLIPR | Future License of Intellectual Property Registry (digital media distribution system) |
What is a cAMP assay?
The assay is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction. The cells are induced with a test compound for an appropriate period of time to modulate cAMP levels.
What is calcium flux assay?
The Calcium Flux assay can be used to monitor the activation of a GPCR via Gq second messenger in a live cell, non-imaging assay format. The active form of the dye becomes fluorescent after binding to intracellular calcium. Additive A is added to prevent the dye from being released out from the cell.
How is calcium influx measured?
There are several directly or indirectly ways to measure intracellular Ca2+ influx:
- Using Ca2+ sensitive dye such as FURa-2 as recommended to you by others.
- using intracellular Ca2+ sensitive microelectrode.
- Performing Whole cell patch clamp recording.
What is a calcium flux assay?
How is cytosolic calcium measured?
Detecting changes in cytosolic calcium is normally performed using a live cell imaging set up with calcium binding dyes that exhibit either an increase in fluorescence intensity or a shift in the emission wavelength upon calcium binding. However, a live cell imaging set up is not freely accessible to all researchers.
How are cAMP levels measured?
The cAMP-Glo™ Max Assay measures cAMP levels through protein kinase A (PKA), which is activated upon release of its regulatory subunits after binding to cAMP (Figure 1). PKA is a component of the cAMP Detection Solution, and once activated, PKA will use any ATP present to phosphorylate its substrate.