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Calcium Chelators / Caged Calcium Chelators
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ALX-450-013 Revised 22-Feb-08
BAPTA . tetrapotassium salt
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SYNONYMS 1,2-bis(2-Aminophenoxy)ethane-N,N,N',N'-tetraacetic acid . 4K
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-450-013-G001   1 g 98.00 USD Add To Cart
Product Specification
FORMULA: C22H20N2O10 . 4K
MW: 472.4 . 156.4
CAS NUMBER: 73630-08-7
PURITY: ≥97% (as anhydride)
APPEARANCE: White to off-white powder.
SOLUBILITY: Soluble in water.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light and moisture.

Product Description
Highly selective Ca2+ chelating agent.
Product Specific Literature References
New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures: R.Y. Tsien; Biochemistry 19, 2396 (1980) Abstract
Calcium-independent increase of transmitter release at frog end-plate by trinitrobenzene sulphonic acid: H. Kijima & T. Tanabe; J. Physiol. 403, 135 (1988) Abstract
Transmitter release at frog end-plate loaded with a Ca2+-chelator, BAPTA: hypertonicity and erythrosin B augment the release independently of internal Ca2+: N. Tanabe & H. Kijima; Neurosci. Lett. 92, 52 (1988) Abstract
Properties of neuroprotective cell-permeant Ca2+ chelators: effects on [Ca2+]i and glutamate neurotoxicity in vitro: M. Tymianski, et al.; J. Neurophysiol. 72, 1973 (1994) Abstract
Chelation of intracellular Ca2+ inhibits murine keratinocyte differentiation in vitro: L. Li, et al.; J. Cell Physiol. 163, 105 (1995) Abstract
 
 
ALX-450-014 Revised 22-Feb-08
BAPTA/AM
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SYNONYMS 1,2-bis(2-Aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-450-014-M010   10 mg 48.00 USD Add To Cart
ALX-450-014-M050   50 mg 195.00 USD Add To Cart
Product Specification
FORMULA: C34H40N2O18
MW: 764.7
CAS NUMBER: 126150-97-8
PURITY: ≥98% (HPLC)
APPEARANCE: White to off-white powder.
SOLUBILITY: Soluble in DMSO (10mg/ml).
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Packaged under inert gas. Protect from light and moisture.
HAZARD: IRRITANT.
IDENTITY: Determined by 1H-NMR.

Product Description
Cell permeable acetoxymethyl ester derivative of BAPTA (Prod. No. ALX-450-013). Useful for controlling intracellular Ca2+ concentration. Induces inactivation of protein kinase C. Also inhibits thapsigargin-induced apoptosis in rat thymocytes.
Product Specific Literature References
Physiological [Ca2+]i level and pump-leak turnover in intact red cells measured using an incorporated Ca chelator: V.L. Lew, et al.; Nature 298, 478 (1982) Abstract
BAPTA induces a decrease of intracellular free calcium and a translocation and inactivation of protein kinase C in macrophages: P. Dieter, et al.; Biol. Chem. Hoppe-Seyler 374, 171 (1993) Abstract
Intracellular calcium chelator, BAPTA-AM, prevents cocaine-induced ventricular fibrillation: G.E. Billman; Am. J. Physiol. 265, H1529 (1993) Abstract
Intracellular Ca2+ signals activate apoptosis in thymocytes: studies using the Ca(2+)-ATPase inhibitor thapsigargin: S. Jiang, et al.; Exp. Cell. Res. 212, 84 (1994) Abstract
Stimulation of the ERK pathway by GTP-loaded Rap1 requires the concomitant activation of Ras, protein kinase C, and protein kinase A in neuronal cells: T. Bouschet, et al.; J. Biol. Chem. 278, 4778 (2003) Abstract; Full Text
 
 
ALX-620-002 Revised 04-Feb-05
FLUO 3 . pentaammonium salt
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SYNONYMS 1-[2-Amino-5-(2,7-dichloro-6-hydroxy-3-oxo-3H-xanthen-9-yl)phenoxy]-2-(2'-amino-5'-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid . 5NH3
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-620-002-M001   1 mg 210.00 USD Add To Cart
Product Specification
FORMULA: C36H30Cl2N2O13 . 5 NH3
MW: 769.6 . 85.2
CAS NUMBER: 134907-84-9
PURITY: ≥75%
APPEARANCE: Reddish brown powder.
SOLUBILITY: Soluble in alkaline solution.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.

Product Description
Ca2+ chelator and indicator. Designed to operate at longer wavelengths of excitation. Among the advantages that it offers are the suitability of argon laser sources for excitations, less interference from endogenous autofluorescence, and the possibility of using other fluorescent probes simultaneously. As with QUIN 2 (Prod. No. ALX-620-006), this chelator responds to Ca2+ without significant changes in excitation or emission maxima hence ratioing techniques cannot be used for the estimation of Ca2+ concentrations. A unique feature of FLUO 3 is that both the acetoxymethyl ester (FLUO 3/AM) and the uncomplexed chelator (FLUO 3) are only weakly fluorescent, but chelation with Ca2+ produces a strong enhancement of fluorescence.
Product Specific Literature References
New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures: R.Y. Tsien; Biochemistry 19, 2396 (1980) Abstract
Fluorescence measurement and photochemical manipulation of cytosolic free calcium: R.Y. Tsien; TINS 11, 419 (1988) Abstract
Kinetics of calcium binding to fluo-3 determined by stopped-flow fluorescence: M. Eberhard & P. Erne; BBRC 163, 309 (1989) Abstract
Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores: A. Minta, et al.; J. Biol. Chem. 264, 8171 (1989) Abstract; Full Text
Photochemically generated cytosolic calcium pulses and their detection by fluo-3: J.P.Y. Kao, et al.; J. Biol. Chem. 264, 8179 (1989) Abstract; Full Text
Use of fluo-3 to measure cytosolic Ca2+ in platelets and neutrophils. Loading cells with the dye, calibration of traces, measurements in the presence of plasma, and buffering of cytosolic Ca2+: J.E. Merritt, et al.; Biochem. J. 269, 513 (1990) Abstract
Improved method for measuring intracellular Ca++ with fluo-3: G.T. Rijkers, et al.; Cytometry 11, 923 (1990) Abstract
Control of mitochondrial matrix calcium: studies using fluo-3 as a fluorescent calcium indicator: A. Saavedra-Molina, et al.; BBRC 167, 148 (1990) Abstract
Effects of photolabile calcium chelators on fluorescent calcium indicators: R.S. Zucker; Cell Calcium 13, 29 (1992) Abstract
 
 
ALX-620-003 Revised 15-Jan-08
FLUO 3/AM
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SYNONYMS 1-[Amino-5-(2,7-dichloro-6-acetomethoxy-3-oxo-3H-xanthen-9-yl)phenoxy]-2-(2'-amino-5'-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid, pentaacetoxy-methyl ester
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-620-003-M001   1 mg 260.00 USD Add To Cart
Product Specification
FORMULA: C51H50Cl2N2O23
MW: 1129.9
CAS NUMBER: 121714-22-5
PURITY: ≥90%
APPEARANCE: Red solid.
SOLUBILITY: Soluble in DMSO (0.9mg/ml) and acetonitrile (2mg/ml).
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.
IDENTITY: Identity determined by 1H-NMR and fluorescence.

Product Description
Cell permeable analog of FLUO 3 (Prod. No. ALX-620-002). Fluorescent probe for Ca2+. After crossing the membrane, the product is quickly metabolized by cytoplasmic esterases to the membrane impermeant FLUO 3.
Product Specific Literature References
Photochemically generated cytosolic calcium pulses and their detection by fluo-3: J.P.Y. Kao, et al.; J. Biol. Chem. 264, 8179 (1989) Abstract; Full Text
Glutamate induces calcium waves in cultured astrocytes: long-range glial signaling: A.H. Cornell-Bell, et al.; Science 247, 470 (1990) Abstract
Control of mitochondrial matrix calcium: studies using fluo-3 as a fluorescent calcium indicator: A. Saavedra-Molina, et al.; BBRC 167, 148 (1990) Abstract
Flow cytometric measurement of cytoplasmic free calcium in human peripheral blood T lymphocytes with fluo-3, a new fluorescent calcium indicator: P.A. Vadenberghe & J.L. Ceuppens; J. Immunol. Meth. 127, 197 (1990) Abstract
Recombinant human tumor necrosis factor alpha induces calcium oscillation and calcium-activated chloride current in human neutrophils. The role of calcium/calmodulin-dependent protein kinase: M.A. Schumann, et al.; J. Biol. Chem. 268, 2134 (1993) Abstract; Full Text
General Information
BACKGROUND/TECHNICAL INFORMATION

General Protocol (for Human T cells)
Reagents:
2mM Fluo 3/AM/DMSO (1mg Fluo 3/AM in 442μl DMSO)
Pluronic F127
Hanks-balanced salt solution (HBSS)
HEPES buffer saline (10mM HEPES, 1mM Na2HPO4, 137mM NaCl, 5mM KCl, 1mM CaCl2, 0.5mM MgCl2, 5mM glucose, 0.1% BSA, pH 7.4)

Protocol:
1. Add 16.5mg Pluronic F127 to Fluo 3/AM/DMSO solution. Pluronic F127 prevents aggregation of Fluo 3/AM in HBSS and helps uptake with cells.
2. Dilute the Fluo 3/AM solution with HBSS to prepare 4 mM Fluo 3/AM working solution.
3. Add the Fluo 3/AM working solution to the cells, and incubate at 37 ºC for 20min.
4. Add 5 volumes of HBSS containing 1% fetal calf serum, and continue the incubation for another 40min.
5. Wash the cells 3 times with HEPES buffer saline. Then resuspend the cells to prepare 1x105 cells/ml solution using HEPES buffer saline.
6. Incubate at 37ºC for 10min. Then use the cells for fluorescent calcium ion detection.
7. Monitor the fluorescence at 528nm (excitation: 490-500nm)

Cell staining conditions differ by cell types, so it is necessary to optimize the conditions for each experiment.



 
 
ALX-620-004 Revised 21-May-08
FURA 2 . pentapotassium salt
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SYNONYMS 1-[2-(5-Carboxyoxazol-2-yl)-6-aminobenzofuran-5-oxy]-2-(2'-amino-5'-methyl-phenoxy)ethane-N,N,N',N'-tetraacetic acid . 5K
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-620-004-M001   1 mg 110.00 USD Add To Cart
Product Specification
FORMULA: C29H22N3O14 . 5K
MW: 636.5 . 195.5
CAS NUMBER: 113694-64-7
PURITY: ≥98% (HPLC)
APPEARANCE: Yellow to orange solid.
SOLUBILITY: Soluble in water.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light and moisture.
IDENTITY:

Identity determined by NMR and fluorescence.


Product Description
Ca2+ chelator that offers more than 30-fold greater fluorescence intensity than QUIN 2, (Prod. No. ALX-620-006), allowing much smaller amounts to be used and minimizes toxic effects and other artifacts. Shows a strong shift in UV absorption maxima on chelation of Ca2+. Ca2+ concentration has been shown to be closely correlated to the ratios of fluorescence, measured at ~510nm, when excitated at 340nm and at 380nm. This ratioing method allows estimation of Ca2+ concentration independent of the dye concentration. Used in many cellular systems, including cardiac myocytes, vascular smooth muscle cells and medullary collecting tubules.
Product Specific Literature References
Spontaneous and experimentally evoked [Ca2+]i-transients in cardiac myocytes measured by means of a fast Fura-2 technique: S. Bals, et al.; Cell Calcium 11, 385 (1990) Abstract
Measurement of intracellular Ca2+ in cultured arterial smooth muscle cells using Fura-2 and digital imaging microscopy: W.F. Goldman, et al.; Cell Calcium 11, 221 (1990) Abstract
Ryanodine reveals multiple contractile and relaxant mechanisms in vascular smooth muscle: simultaneous measurements of mechanical activity and of cytoplasmic free Ca2+ level with fura-2: T. Hisayama, et al.; Br. J. Pharmacol. 100, 677 (1990) Abstract
Cholinergic agonists increase cell calcium in rat medullary collecting tubules. A fura-2 study: J. Marchetti, et al.; Pfluegers Arch. 416, 561 (1990) Abstract
Estimation of intracellular calcium activity in confluent monolayers of primary cultures of quail medullary bone osteoclasts: S. Arkle, et al.; Exp. Physiol. 79, 975 (1994) Abstract
General Information
BACKGROUND/TECHNICAL INFORMATION FURA 2 pentapotassium salt is not cell-permeable. Cells need to be loaded by pressure microinjection or permeabilized prior to use this compound. These approaches are only suggestions as optimal conditions should be determined by individual users.
 
 
ALX-620-005 Revised 22-Feb-08
FURA 2/AM
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SYNONYMS 1-[2-(5-Carboxyoxazol-2-yl)-6-aminobenzofuran-5-oxy]-2-(2'-amino-5'-methyl-phenoxy)ethane-N,N,N',N'-tetraacetic acid, pentaacetoxymethyl ester
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-620-005-M001   1 mg 135.00 USD Add To Cart
Product Specification
FORMULA: C44H47N3O24
MW: 1001.9
CAS NUMBER: 108964-32-5
PURITY: ≥98% (HPLC)
APPEARANCE: Off-white to yellow solid.
SOLUBILITY: Soluble in acetonitrile or DMSO.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light and moisture.
IDENTITY: Determined by 1H-NMR and fluorescence.

Product Description
Cell permeable analog of FURA 2 (Prod. No. ALX-620-004). Fluorescent probe for Ca2+. After crossing the membrane, the product is quickly metabolized by cytoplasmic esterases to the membrane impermeant FURA 2.
Product Specific Literature References
Fura-2 measurement of cytosolic free Ca2+ in monolayers and suspensions of various types of animal cells: A. Malgaroli, et al.; J. Cell. Biol. 105, 2145 (1987) Abstract
A Ca2+-insensitive form of fura-2 associated with polymorphonuclear leukocytes. Assessment and accurate Ca2+ measurement: M. Scanlon, et al.; J. Biol. Chem. 262, 6308 (1987) Abstract; Full Text
Intracellular calibration of the fluorescent calcium indicator Fura-2: D.A. Williams & F.S. Fay; Cell Calcium 11, 75 (1990) Abstract
Stretch-induced increases in intracellular calcium of isolated vascular smooth muscle cells: M.J. Davis, et al.; Am. J. Physiol. 263, H1292 (1992) Abstract
Positive and negative inotropic effects of phorbol 12-myristate 13- acetate: relationship to PKC-dependence and changes in [Ca2+]i: C.A. Ward & M.P. Moffat; J. Mol. Cell. Cardiol. 24, 937 (1992) Abstract
 
 
ALX-450-004 Revised 04-Feb-05
INDO 1 . pentapotassium salt
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SYNONYMS 1-[2-Amino-5-(6-carboxyindol-2-yl)phenoxy]-2-(2'-amino-5'-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid . 5K
PRODUCT LINE Other Products
PRODUCT CATEGORY Dyes / Stains / Fluorescent Probes / Fluorescent Labels
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ALX-450-004-M001   1 mg 95.00 USD Add To Cart
Product Specification
FORMULA: C32H26N3O12 . 5K
MW: 644.6 . 195.5
CAS NUMBER: 132319-56-3
PURITY: ≥97%
APPEARANCE: Off-white solid.
SOLUBILITY: Soluble in water.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.

Product Description
Ca2+ chelator and indicator of intracellular and extracellular Ca2+.  Particularly useful in flow cytometry studies.
Product Specific Literature References
New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures: R.Y. Tsien; Biochemistry 19, 2396 (1980) Abstract
A new generation of Ca2+ indicators with greatly improved fluorescence properties: G. Grynkiewicz, et al.; J. Biol. Chem. 260, 3340 (1985) Abstract; Full Text
Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry: P.S. Rabinovitch, et al.; J. Immunol. 137, 952 (1986) Abstract
CD5 antibodies increase intracellular ionized calcium concentration in T cells: C.H. June, et al.; J. Immunol. 138, 2782 (1987) Abstract
Diminished calcium influx in lectin-stimulated T cells from old mice: R.A. Miller, et al.; J. Cell. Physiol. 132, 337 (1987) Abstract
Calcium mobilization in human platelets using indo-1 and flow cytometry: L.K. Jennings, et al.; Blood 74, 2674 (1989) Abstract
Analysis of cytosolic ionized calcium variation in polymorphonuclear leukocytes using flow cytometry and Indo-1 AM: M. Lopez, et al.; Cytometry 10, 165 (1989) Abstract
Cytosolic and vacuolar Ca2+ concentrations in yeast cells measured with the Ca2+-sensitive fluorescence dye indo-1: D. Halachmi & Y. Eilam; FEBS Lett. 256, 55 (1989) Abstract
Intracellular Ca2+ measurement with Indo-1 in substrate-attached cells: advantages and special considerations: M. Wahl, et al.; Cell Calcium 11, 487 (1990) Abstract
Calcium changes in immune complex-stimulated human neutrophils. Simultaneous measurement of receptor occupancy and activation reveals full population stimulus binding but subpopulation activation: B.A. Brunkhorst, et al.; J. Biol. Chem. 266, 13035 (1991) Abstract; Full Text
 
 
ALX-450-005 Revised 04-Feb-05
INDO 1/AM
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SYNONYMS 1-[2-Amino-5-(6-carboxyindol-2-yl)phenoxy]-2-(2'-amino-5'-methylphenoxy)ethane-N,N,N',N'-tetraacetic acid, pentaacetoxymethylester
PRODUCT LINE Other Products
PRODUCT CATEGORY Dyes / Stains / Fluorescent Probes / Fluorescent Labels
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ALX-450-005-M001   1 mg 130.00 USD Add To Cart
ALX-450-005-M005   5 mg 520.00 USD Add To Cart
Product Specification
FORMULA: C47H51N3O22
MW: 1009.9
CAS NUMBER: 112926-02-0
PURITY: ≥97%
APPEARANCE: Off-white solid.
SOLUBILITY: Soluble in DMSO.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.

Product Description
Cell permeable analog of INDO 1 (Prod. No. ALX-450-004). Fluorescent probe for Ca2+. After crossing the membrane, the product is rapidly hydrolyzed by cytoplasmic esterases to the membrane impermeant INDO 1.
Product Specific Literature References
New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures: R.Y. Tsien; Biochemistry 19, 2396 (1980) Abstract
A new generation of Ca2+ indicators with greatly improved fluorescence properties: G. Grynkiewicz, et al.; J. Biol. Chem. 260, 3440 (1985) Abstract; Full Text
Measuring cytosolic free calcium concentration in endothelial cells with indo-1: the pitfall of using the ratio of two fluorescence intensities recorded at different wavelengths: A. Luckhoff; Cell Calcium 7, 233 (1986) Abstract
Cross-linking of surface immunoglobulin on B lymphocytes induces both intracellular Ca2+ release and Ca2+ influx: analysis with indo-1: M.K. Bijsterbosch, et al.; BBRC 137, 500 (1986) Abstract
Heterogeneity among T cells in intracellular free calcium responses after mitogen stimulation with PHA or anti-CD3. Simultaneous use of indo-1 and immunofluorescence with flow cytometry: P.S. Rabinovitch, et al.; J. Immunol. 137, 952 (1986) Abstract
Cytosolic and vacuolar Ca2+ concentrations in yeast cells measured with the Ca2+-sensitive fluorescence dye indo-1: D. Halachmi & Y. Eilam; FEBS Lett. 256, 55 (1989) Abstract
Calcium mobilization in human platelets using indo-1 and flow cytometry: L.K. Jennings, et al.; Blood 74, 2674 (1989) Abstract
Analysis of cytosolic ionized calcium variation in polymorphonuclear leukocytes using flow cytometry and Indo-1 AM: M. Lopez, et al.; Cytometry 10, 165 (1989) Abstract
Intracellular Ca2+ measurement with Indo-1 in substrate-attached cells: advantages and special considerations: M. Wahl, et al.; Cell Calcium 11, 487 (1990) Abstract
Calcium homeostasis in dissociated embryonic neurons: a flow cytometric analysis: J.P. Grierson, et al.; J. Neurophysiol. 67, 704 (1992) Abstract
 
 
ALX-620-006 Revised 04-Mar-05
QUIN 2 . tetrapotassium salt
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SYNONYMS 2-[(2-bis-[Carboxymethyl]amino-5-methylphenoxy)-methyl]-6-methoxy-8-bis[carboxymethyl]aminoquinoline . 4K
PRODUCT LINE Signal Transduction
PRODUCT CATEGORY Calcium Chelators / Caged Calcium Chelators
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ALX-620-006-M005   5 mg 35.00 USD Add To Cart
Product Specification
FORMULA: C26H23N3O10 . 4K
MW: 537.5 . 156.4
CAS NUMBER: 73630-23-6
MERCK INDEX: 14: 8042
PURITY: ≥95% (contains 11-13% water)
APPEARANCE: Yellow powder.
SOLUBILITY: Soluble in water.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: Protect from light.