Indo-1 (sodiumsalt) is a ratiometric fluorescent calcium indicator. It is ideal for analyses using flow cytometry, as it uses a single excitation source, typically 349-364 nm light from an argon-ion laser. The emission maximum shifts from 475-485 nm without calcium to 400-410 nm when Indo-1 (sodiumsalt) binds calcium. Indo-1 (sodiumsalt) is prone to photobleaching, which limits its usefulness in methods involving microscopy.
XTT is a cell-impermeable, negatively charged tetrazolium dye that produces a water-soluble formazan when reduced at the cell surface by cellular-derived NADH and an electron mediator.1,2It is frequently used in colorimetric assays to measure cell proliferation, cytotoxicity, and apoptosis.3 1.Berridge, M.V., Tan, A.S., and Herst, P.M.Tetrazolium dyes as tools in cell biology: New insights into their cellular reductionBiotechnol. Ann. Rev.11127-152(2005) 2.Berridge, M.V., and Tan, A.S.Trans-plasma membrane electron transport: A cellular assay for NADH-and NADPH-oxidase based on extracellular, superoxide-mediated reduction of the sulfonated tetazolium salt WST-1Protoplasma20574-82(1998) 3.Sutherland, M.W., and Learmonth, B.A.The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutaseFree Radical Research27(3)283-289(1997)
Fura-2 is a ratiometric fluorescent calcium indicator that can be used to detect calcium in cells. It is a pentacarboxylate that displays excitation maxima of 340 and 380 nm at high and low calcium concentrations, respectively, when the emission is fixed at 510 nm, enabling determination of ratiometric measurements of calcium influx in live cells.
Fluo-3 is a fluorescent calcium indicator commonly used in flow cytometry and cell-based experiments to detect changes in intracellular calcium levels. Its absorption maximum at 506 nm makes it compatible with excitation at 488 nm by argon-ion laser sources. Fluo-3 provides intense fluorescence upon binding calcium, detected at a maximum emission at 526 nm which can be monitored by FL1 (green, 525 nm band pass) sensors in flow cytometry.
4-Methylumbelliferyl 2-sulfamino-2-deoxy-α-D-Glucopyranoside (4-MU-α-GlcNS) is a fluorogenic substrate of heparin sulphamidase. Heparin sulphamidase cleaves 4-MU-α-GlcNS to yield 4-MU-α-GlcNH2, which is then cleaved by α-glucosaminidase to release the fluorescent product 4-MU, which displays an emission maxima of 445-454 nm. The excitation maxima for 4-MU is pH-dependent: 330, 370, and 385 nm at pH 4.6, 7.4, and 10.4, respectively. 4-MU-α-GlcNS has been used to quantify heparin sulphamidase deficiencies associated with Mucopolisaccaridosis IIIA and other lysosomal disorders.
4-Methylumbelliferyl β-D-Galactopyranoside-6-sulfate (sodiumsalt) (4-MU-Gal-6S) is a fluorogenic substrate used to quantify N-acetylgalactosamine-6-sulphatase (GALNS) activity. 4-MU-Gal-6S is cleaved by GALNS to release the fluorescent moiety 4-MU. 4-MU fluorescence is pH-dependent with excitation maxima of 320 and 360 nm at low (1.97-6.72) and high (7.12-10.3) pH, respectively, and an emission maximum ranging from 445 to 455 nM, increasing as pH decreases. It has been used to detect Morquio disease type A, a lysosomal storage disorder in which GALNS is deficient. 4-MU-Gal-6S can be used to assess GALNS activity in a very small blood volume to determine the extent of deficiency.
Rhod-2 (sodiumsalt) is a water-soluble, red fluorescent calcium indicator. It exhibits a significant shift in fluorescence intensity upon calcium binding (ex max = 549 nm; calcium-free v. ex/em max = 552/581 nm; calcium-bound). [1][2] Unlike the UV-excitable indicators fura-2 and indo-1 , there is no accompanying spectral shift.