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ALX-620-054

Revised 11-Oct-06

4AF DA
SYNONYMS	4-Aminofluorescein diacetate
PRODUCT LINE	Nitric Oxide Pathway
PRODUCT CATEGORY	Nitric Oxide & NOS Detection / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-620-054-M001		1 mg	136.00 USD

Product Specification

FORMULA:	C₂₄H₁₇NO₇
MW:	431.4
PURITY:	≥99% (HPLC)
APPEARANCE:	White to off-white powder.
SOLUBILITY:	Soluble in DMSO.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	+4°C
HANDLING:	Protect from light. Keep under inert gas.

Product Description

Negative control compound for the fluorescent probes DAF-2 (Prod. No. ALX-620-052) and DAF-2 DA (Prod. No. ALX-620-056).

Product Specific Literature References

Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA: H. Kojima, et al.; Neuroreport 9, 3345 (1998) Abstract

Further Categories Containing This Product:

Dyes / Stains / Fluorescent Probes / Fluorescent Labels

ALX-620-029

Revised 05-May-08

ABD-F
SYNONYMS	4-Fluoro-7-sulfoamoylbenzofurazan
PRODUCT LINE	Other Products
PRODUCT CATEGORY	Dyes / Stains / Fluorescent Probes / Fluorescent Labels

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-620-029-M010		10 mg	95.00 USD
ALX-620-029-M050		50 mg	320.00 USD

Product Specification

FORMULA:	C₆H₄FN₃O₃S
MW:	217.2
CAS NUMBER:	91366-65-3
PURITY:	≥99%
APPEARANCE:	White to yellow powder.
SOLUBILITY:	Soluble in acetonitrile.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light and moisture.

Product Description

Highly reactive fluorogenic reagent for labelling thiol compounds. Selectively labels protein cysteine residues without side chain modification or N-terminal blockage. Potent vasorelaxant.

Product Specific Literature References

New Fluorogenic Reagent Having Halogenobenzofurazan Structure for Thiols: 4-(Aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole: T. Toyo'oka and K. Imai; Anal. Chem. 56, 2461 (1984)
Isolation and characterization of cysteine-containing regions of proteins using 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole and high- performance liquid chromatography: T. Toyo'oka and K. Imai; Anal. Chem. 57, 1931 (1985)
Amino acid composition analysis of minute amounts of cysteine- containing proteins using 4-(aminosulfonyl)-7-fluoro-2,1,3- benzoxadiazole and 4-fluoro-7-nitro-2,1,3-benzoxadiazole in combination with HPLC: T. Toyo'oka, et al.; Biomed. Chromatogr. 1, 15 (1986) Abstract
Fluorometric assay of thiols with fluorobenzoxadiazoles: K. Imai and T. Toyo'oka; Meth. Enzymol. 143, 67 (1987) Abstract

ALX-380-283

Revised 21-May-08

7-Amino-actinomycin D
SYNONYMS	7-AAD 7-Aminoactinomycin C₁
PRODUCT LINE	Natural Products / Antibiotics
PRODUCT CATEGORY	Antitumor Antibiotics

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-380-283-M001		1 mg	90.00 USD
ALX-380-283-M005		5 mg	360.00 USD

Product Specification

FORMULA:	C₆₂H₈₇N₁₃O₁₆
MW:	1270.4
CAS NUMBER:	7240-37-1
RTECS:	AU1579000
SOURCE/HOST:	Semisynthetic from actinomycin D (Prod. No. ALX-380-009).
PURITY:	≥96% (HPLC)
APPEARANCE:	Red to dark purple powder.
SOLUBILITY:	Soluble in DMSO (20mg/ml), methanol or 100% ethanol; slightly soluble in water.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light. Hygroscopic.
HAZARD:	VERY TOXIC.

Product Description

Membrane impermeable fluorescent DNA intercalator. Used as a fluorescent DNA stain. Inhibitor of DNA-primed RNA polymerase and DNA polymerase. Induces apoptosis and is used as a tool to identify apoptotic cells. Cytochemical probe. Exhibits growth-inhibitory activity against certain leukemias and sarcomas. Has antibacterial properties.

Product Specific Literature References

7-Amino-actinomycin D as a cytochemical probe. I. Spectral properties: J.E. Gill, et al.; J. Histochem. Cytochem. 23, 793 (1975) Abstract; Full Text
7-substituted actinomycin D analogs. Chemical and growth-inhibitory studies: S.K. Sengupta, et al.; J. Med. Chem. 18, 1175 (1975) Abstract
Simultaneous cell cycle analysis and two-color surface immunofluorescence using 7-amino-actinomycin D and single laser excitation: applications to study of cell activation and the cell cycle of murine Ly-1 B cells: P.S. Rabinovitch, et al.; J. Immunol. 136, 2769 (1986) Abstract
Binding specificities of actinomycin D to non-self-complementary -XGCY-tetranucleotide sequences: F.M. Chen; Biochemistry 31, 6223 (1992) Abstract
Dead cell discrimination with 7-amino-actinomycin D in combination with dual color immunofluorescence in single laser flow cytometry: I. Schmid, et al.; Cytometry 13, 204 (1992) Abstract
Improved staining method for the simultaneous flow cytofluorometric analysis of DNA content, S-phase fraction, and surface phenotype using single laser instrumentation: K. Toba, et al.; Cytometry 13, 60 (1992) Abstract
Flow cytometric immunofluorescence assay for quantification of cyclobutyldithymine dimers in separate phases of the cell cycle: R.J. Berg, et al.; Carcinogenesis 14, 103 (1993) Abstract
Simultaneous three-color analysis of the surface phenotype and DNA-RNA quantitation using 7-amino-actinomycin D and pyronin Y: K. Toba, et al.; J. Immunol. Methods 182, 193 (1995) Abstract
Hematopoietic cell protein-tyrosine phosphatase-deficient motheaten mice exhibit T cell apoptosis defect: X. Su, et al.; J. Immunol. 156, 4198 (1996) Abstract
The use of 7-amino actinomycin D in identifying apoptosis: simplicity of use and broad spectrum of application compared with other techniques: N.J. Philpott, et al.; Blood 87, 2244 (1996) Abstract; Full Text
A rapid method for measuring apoptosis and dual-color immunofluorescence by single laser flow cytometry: I. Schmid, et al.; J. Immunol. Methods 170, 145 (1996) Abstract
Strategies for phenotyping apoptotic peripheral human lymphocytes comparing ISNT, annexin-V and 7-AAD cytofluorometric staining methods: H. Lecoeur, et al.; J. Immunol. Methods 209, 111 (1997) Abstract
Cell kinetic study of normal human bone marrow hematopoiesis and acute leukemia using 7AAD/PY: K. Toba, et al.; Eur. J. Haematol. 64, 10 (2000) Abstract
Live-cell assay for detection of apoptosis by dual-laser flow cytometry using Hoechst 33342 and 7-amino-actinomycin D: I. Schmid, et al.; Nat. Protoc. 2, 187 (2007) Abstract

Related Products

ALX-380-009

Actinomycin D

Further Categories Containing This Product:

Antibiotics - Apoptosis Inducers & Inhibitors • Transcription Inhibitors / Related Products • Antitumor Agents (Apoptosis Inducers) • Apoptosis & Necrosis Detection Other Products • DNA Intercalators & Crosslinkers • Dyes / Stains / Fluorescent Probes / Fluorescent Labels

ALX-610-028

Revised 17-Jan-06

7-Amino-4-methylcoumarin
SYNONYMS	AMC Coumarin 120
PRODUCT LINE	Other Products
PRODUCT CATEGORY	Dyes / Stains / Fluorescent Probes / Fluorescent Labels

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-610-028-M010		10 mg	20.00 USD

Product Specification

FORMULA:	C₁₀H₉NO₂
MW:	175.2
CAS NUMBER:	26093-31-2
PURITY:	≥98%
APPEARANCE:	Off-white to yellow powder.
SOLUBILITY:	Soluble in dimethylformamide, acetone or DMSO.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	+4°C
HANDLING:	Keep cool and dry. Keep under inert gas.

Product Description

Fluorogenic chromophore. Ex(max): 380nm; Em(max) 460nm. Reference compound.

ALX-610-027

Revised 29-Nov-04

7-Amino-4-trifluoromethylcoumarin
SYNONYMS	AFC Coumarin 151
PRODUCT LINE	Other Products
PRODUCT CATEGORY	Dyes / Stains / Fluorescent Probes / Fluorescent Labels

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-610-027-M010		10 mg	15.00 USD

Product Specification

FORMULA:	C₁₀H₆F₃NO₂
MW:	229.2
CAS NUMBER:	53518-15-3
PURITY:	≥97%
APPEARANCE:	Off white to yellow powder.
SOLUBILITY:	Soluble in DMSO, DMF or methanol.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	+4°C
HANDLING:	Protect from light. Keep under inert gas.

Product Description

Fluorogenic chromophore. Ex(max): 400nm; Em(max) 505nm. Reference compound.

ALX-620-075

Revised 25-Sep-08

APF
SYNONYMS	Aminophenyl fluorescein 2-[6-(4'-Amino)phenoxy-3H-xanthen-3-on-9-yl]benzoic acid
PRODUCT LINE	Oxidative Stress
PRODUCT CATEGORY	Oxidative Stress Markers & Reagents / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-620-075-M001		1 mg	430.00 USD

Product Specification

FORMULA:	C₂₆H₁₇NO₅
MW:	423.4
CAS NUMBER:	359010-70-1
CONCENTRATION:	~5mM
PURITY:	≥98% (HPLC)
FORMULATION:	Liquid. In 0.47ml dimethyl formamide.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	+4°C
USE/STABILITY:	Prepare 500-5’000-fold dilution (~10-1µM) in phosphate buffer (0.1M phosphate, pH 7.4) immediately before use. BSA, phenol red and amines may affect the fluorescence and must be used with caution. Do not store the dilutions.
HANDLING:	Keep under inert gas. Protect from light. After opening, prepare aliquots and store at +4°C.
HAZARD:	HARMFUL.

Product Images

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Product Description

Fluorescent reagent (Ex(max): 490nm; Em(max): 515nm) for the detection of highly reactive oxygen species (hROS). Immediately reacts with hROS such as hydroxyl radical, peroxynitrite and hypochlorite, and the fluorescence intensity greatly increases. Use of APF together with HPF (Prod. No. ALX-620-074) also allows for specific detection of hypochlorite (^-OCl) to elucidate reliable the roles of ^-OCl in biological systems such as neutrophils. In addition, peroxynitrite can be detected in distinction from nitric oxide and superoxide since APF does not react with nitric oxide, superoxide and hydrogen peroxide. Moreover, APF is resistant to light-induced autooxidation. Not for sale in Japan.

Product Specific Literature References

Development of novel fluorescence probes that can reliably detect reactive oxygen species and distinguish specific species: K. Setsukinai, et al.; J. Biol. Chem. 278, 3170 (2003) Abstract

General Information

BACKGROUND/TECHNICAL INFORMATION

Precautions for use:
1. Prepare the diluted solution just before use and use it up.
2. The dilution buffer should be adjusted to pH 7.0-7.5. Please note that bovine serum albumin (BSA) and phenol red might interfere with the fluorescence measurement.

Light-induced autoxidation (Figure 3)
After adding HPF or DCFH-DA (10µM), the cells were incubated for 30 min at 37°C in the dark. The fluorescence images were obtained with a confocal fluorescent microscope (excitation wavelength: 488nm, fluorescence emission wavelength: using a 515nm barrier filter). The cells were laser-irradiated again at 488nm for 10 sec and fluorescence images were obtained again (see Figure 3).

Fluorescence images of HPF- or APF-loaded neutrophils (Figure 4)
Neutrophils were obtained from porcine blood and suspended in Krebs-Ringer phosphate buffer (114mM NaCI, 4.6mM KCI, 2.4mM MgSO₄, 1.0mM CaCI₂, 15mM NaH₂PO₄/Na₂HPO₄, pH 7.4) and kept on ice until use. Then they were seeded onto a glass-bottomed dish. The cells were loaded with HPF or APF (10µM) by incubation for 30min at room temperature and then stimulated with PMA (including 2ng/ml; 0.1% DMF as a cosolvent). Fluorescence images were obtained with a confocal fluorescent microscope before and 10 min after the stimulation with PMA (excitation wavelength: 488nm, fluorescence emission wavelength: 505-550nm; using a 505-550nm barrier filter).

Detection of hROS in the HRP/H₂O₂system using HPF and APF (Figure 5)
Reaction timecourse
Fluorescence probe reagents (final 10µM; 0.1% DMF as a cosolvent) were added to sodium phosphate buffer (0.1M; pH 7.4) containing HRP (0.2µM). H₂O₂ (final 1µM) was added at the time indicated by the arrow. The fluorescence intensities were measured at excitation wavelength of 490nm and fluorescence emission wavelength of 515nm. Relation between the amount of added H₂O₂ and fluorescence increase in the HRP/H₂O₂ system.

Relation between the amount of added H₂O₂ and fluorescence increase in the HRP/H₂O₂ system
Fluorescence probe reagents (final 10µM; 0.1% DMF as a cosolvent) were added to sodium phosphate buffer (0.1M; pH 7.4) containing HRP (0.2µM). The fluorescence intensities were measured at excitation wavelength of 490nm and fluorescence emission wavelength of 515nm.

Related Products

ALX-620-074

HPF

Further Categories Containing This Product:

Peroxynitrite / Scavengers / Detection • Dyes / Stains / Fluorescent Probes / Fluorescent Labels

ALX-450-013

Revised 22-Feb-08

BAPTA . tetrapotassium salt
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

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-450-013-G001		1 g	98.00 USD

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

Further Categories Containing This Product:

Nuclear Envelope & Nuclear Transport • Dyes / Stains / Fluorescent Probes / Fluorescent Labels

ALX-620-051

Revised 30-Jun-08

Bisbenzimide H33258 . trihydrochloride
SYNONYMS	HOE 33258 2'-4-(Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bis-1H-benzimidazole . 3HCl
PRODUCT LINE	Other Products
PRODUCT CATEGORY	Dyes / Stains / Fluorescent Probes / Fluorescent Labels

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-620-051-M010		10 mg	22.00 USD
ALX-620-051-M050		50 mg	35.00 USD

Product Specification

FORMULA:	C₂₅H₂₄N₆O . 3HCl
MW:	424.4 . 109.4
CAS NUMBER:	23491-45-4
RTECS:	SM1140500
PURITY:	≥98% (TLC)
APPEARANCE:	Yellow to greenish powder.
SOLUBILITY:	Soluble in water (10mg/ml).
SHIPPING:	AMBIENT
LONG TERM STORAGE:	-20°C
HANDLING:	Protect from light.
HAZARD:	MAY BE MUTAGENIC. HARMFUL.
IDENTITY:	Identity determined by IR and ¹H-NMR.

Product Description

Cell permeable fluorescent dye with low cytotoxicity. Useful for staining DNA, chromosomes and nuclei [1,2]. Can be used with chromomycin A₃ (Prod. No. ALX-380-055) to distinguish chromosomes by their total DNA concentration and their DNA base composition [3,4]. Binds to A:T pairs in DNA. Also useful for controlling gene expression through binding to RNA aptamers inserted into 5' untranslated region of a messenger RNA [5]. Increases the rate of channel-mediated Ca²⁺ efflux from junctional cytoplasmic reticulum vesicles [6]. Ex(max): 346nm; Em(max): 460nm.

Product Specific Literature References

[1] Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis: S.A. Latt & G. Stetten; J. Histochem. Cytochem. 24, 24 (1976) Abstract
[2] Comparison of cellular and nuclear flow cytometric techniques for discriminating apoptotic subpopulations: K.H. Elstein & R.M. Zucker; Exp. Cell. Res. 211, 322 (1994) Abstract
[3] Bacterial characterization by flow cytometry: M.A. Van Dilla, et al.; Science 220, 620 (1983) Abstract
[4] Determination of guanine-plus-cytosine content of bacterial DNA by dual-laser flow cytometry: C.A. Sanders, et al.; J. Gen. Microbiol. 136, 359 (1990) Abstract
[5] Controlling gene expression in living cells through small molecule-RNA interactions: G. Werstuck & M.R. Green; Science 282, 296 (1998) Abstract
[6] Activation and inhibition of the sarcoplasmic reticulum Ca2+ channel by the polycationic dyes Hoechst 33342 and Hoechst 33258: T.J. Beeler & K. Gable; J. Membr. Biol. 135, 109 (1993) Abstract

FORMULA:	C₂₇H₂₈N₆O . 3HCl
MW:	452.6 . 109.4
CAS NUMBER:	23491-52-3
PURITY:	≥95%
APPEARANCE:	Yellow to green powder.
SOLUBILITY:	Soluble in water.
SHIPPING:	AMBIENT
LONG TERM STORAGE:	+4°C
HANDLING:	Protect from light.
HAZARD:	TOXIC. MAY BE CARCINOGENIC.