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Key Documents

QIA39

Sigma-Aldrich

FragEL DNA Fragmentation Detection Kit, Fluorescent - TdT Enzyme

Synonym(s):

TUNEL Assay

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About This Item

UNSPSC Code:
41116133
NACRES:
NA.51

usage

sufficient for 50 tests

Quality Level

species reactivity (predicted by homology)

all

manufacturer/tradename

Calbiochem®

storage condition

OK to freeze
avoid repeated freeze/thaw cycles

technique(s)

flow cytometry: suitable

input

sample type cell suspension(s)
sample type frozen section(s)
sample type paraffin section(s)
sample type fixed-cell preparation(s)

detection method

fluorometric

shipped in

wet ice

storage temp.

−20°C

General description

Allows for detection of apoptosis at the individual cell level. Apoptosis morphology is easily detected. Detection is by flow cytometry or fluorescence microscopy. The kit mounting media permits the observance of the total cell population.
Cat. No. QIA39 is a fluorescein-conjugated version of our widely referenced TdT Colorimetric FragEL DNA Fragmentation Detection Kit (Cat. No. QIA33) for non-isotopic labeling. Terminal deoxynucleotidyl transferase (TdT) binds to exposed 3′-OH ends of DNA fragments generated in response to apoptotic signals and catalyzes the addition of fluorescein-labeled and unlabeled deoxynucleotides. When excited, fluorescein generates an intense signal which can be detected by fluorescence microscopy or flow cytometry. The mounting media sustains the fluorescent signal from samples labeled on slides and, therefore, aids in the morphological evaluation and characterization of normal and apoptotic cells. Non-apoptotic cells can be visualized by a DAPI filter.

Components

Proteinase K, Equilibration and Labeling Buffers, TdT Enzyme (Cat. No. PF060), Control Slides, Mounting Media (Cat. No. HC08), and a user protocol.

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)

Specifications

Assay Time: 2 h

Principle

The Calbiochem Fluorescein-FragEL DNA Fragmentation Detection Kit is a non-isotopic system for the labeling of DNA breaks in apoptotic nuclei in paraffin-embedded tissue sections, tissue cryosections, in cell preparations fixed on slides, or in cell suspensions. Detection and analysis of labeling is possible by flow cytometry or fluorescence microscopy.

Storage and Stability

Fluorescein-FragEL kit components are shipped on dry ice. Upon receipt, store kit at -20°C in a non-frost-free freezer. To avoid reagent loss in tube caps, briefly pulse spin all thawed solutions before removing caps. (see General Considerations below).

Other Notes

Darzynkiewicz, Z., et al. 1997. Cytometry27, 1.
Frey, T. 1997. Cytometry28, 253.
Shapiro, H.M. 1995. Practical Flow Cytometry, Third Edition. Wiley-Liss Inc., New York, New York.
Darzynkiewicz, Z., et al. 1992. Cytometry13, 795.
Gavrieli, Y., et al. 1992. J. Cell Biol.119, 493.
Fawthrop, D. J., et al. 1991. Arch. Toxicol.65, 437.
Martin, S.J., et al. 1990. J. Immunology145, 1859.
Wyllie, A. H. 1980. Nature284, 555.
Kerr, J. F. R., et al. 1972. Br. J. Cancer26, 239.
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Parafilm is a registered trademark of Bemis Company, Inc.

Signal Word

Danger

Hazard Classifications

Acute Tox. 4 Inhalation - Acute Tox. 4 Oral - Aquatic Chronic 2 - Carc. 1B - Eye Irrit. 2

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects


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