Fluorescent in vivo Detector for intracellular Ca Ions

Case12 - measure intracellular Ca2+ changes in vivo

• High dynamic range calcium indicator
• Direct expression in cells
• No cofactors, substrate addition, or chemical staining required
• High selectivity and sensitivity
• High brightness of fluorescent response
• Fast maturation at 37°C
• Relatively high pH stability

Calcium ions—atoms of calcium with a +2 charge—travel in and out of cells through pores in the cell membrane called ion channels. Once inside the cell, calcium ions trigger a cascade of biochemical reactions that lead to genes being turned on and off. Activated genes produce proteins, which carry out all of life´s functions.
Calcium, for example, is key to making the connections between neurons stronger and increasing the stability of the "wiring" within the brain. Malfunctions of calcium signaling are involved in neurological disorders such as Alzheimer Disease, Parkinson Disease, Multiple Sclerosis and others.

Case12 is a high dynamic range genetically encoded fluorescent sensor for detection of intracellular Ca2+ changes (Souslova et al.). The sensor allows direct measurement of changes of calcium concentration in a physiological range from hundred nanomoles to micromoles with a high signal-to-noise ratio. Binding of Ca2+ is fast and reversible, allowing monitoring high-frequency Ca2+ oscillations. In response to Ca2+ concentration rise, Case12 shows up to 12-fold increase of fluorescence brightness. Fluorescence of Case12 is characterized by single excitation/emission maxima peaked at 491/516 nm.

Available variants and fusions
Case12 codon usage is optimized for high expression in mammalian cells, but it can be successfully expressed in other heterological systems.

Case12-Cyto When expressed in mammalian cells, this variant is evenly distributed throughout the cytosol and the nucleus.

Case12-Mem Membrane localization signal of neuromodulin is fused to Case12 N-terminus. The signal provides for posttranslational palmitoylation that targets Case12 to cellular membranes.

Case12-Mito Mitochondrial targeting sequence derived from subunit VIII precursor of human cytochrome C oxidase is fused to Case12 N-terminus. When expressed in mammalian cells, this variant is localized in mitochondria.

Gateway® Entry Vectors The Case12-cyto or mito coding sequence is flanked by attL1 and attL2 sites allowing easy site-specific recombination. This provides a rapid and highly efficient way to transfer the Case12-cyto or -mito gene into a number of Gateway® destination vectors for expression in different experimental systems.

Case12 Details
Cell Lines stably transfected with Case12

Description	Cat#	Size	Price
Case12-Cyto Vector	FP991-EV	20 ug	400 €	DETAILS
Case12-Mem Vector	FP993-EV	20 ug	400 €	DETAILS
Case12-Mito Vector	FP992-EV	20 ug	400 €	DETAILS
Gateway Case12-cyto entry vector	FP994-EV	20 ug	400 €	DETAILS
Gateway Case12-mito entry vector	FP995-EV	20 ug	400 €	DETAILS