Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) orders@cellsignal.com

Support: 877-678-TECH (8324)

Web: info@cellsignal.com cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

IHC-P, IF-IC, FC-FP

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Product Information

Storage

All components in this kit are stable for at least 12 months when stored at the recommended temperature.

Specificity / Sensitivity

Tissue processing or cell degradation may result in labeling of nuclei in non-apoptotic cells.

Product Description

TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) is a method by which a fluorophore conjugated nucleotide is enzymatically linked to the 3′ end of fragmented DNA. Since DNA fragmentation is a hallmark of apoptosis, the TUNEL assay has become a well established method to monitor apoptosis in situ.

The TUNEL Assay Kit (Fluorescence, 594 nm) provides you with the buffers, enzyme, and dUTP fluorophore conjugate necessary to complete the TUNEL reaction in fixed cells or tissue. This kit is intended for use with fluorescence microscopy and/or flow cytometry.

Background

Apoptosis is a regulated cellular suicide mechanism characterized by nuclear condensation, cell shrinkage, membrane blebbing, and DNA fragmentation (1). During late stages of apoptosis, DNA is fragmented by an endonuclease that cleaves the chromatin into nucleosomal units which can be visualized on gels as DNA laddering (2). DNA fragmentation also serves as a basis for monitoring apoptosis in situ using the TUNEL assay (3). In this assay, terminal deoxynucleotidyl transferase enzymatically incorporates a fluorophore conjugated nucleotide to the 3' end of fragmented DNA. Apoptosis can be monitored by an increase in TUNEL staining within intact cells.

  1. Kerr, J.F. et al. (1972) Br J Cancer 26, 239-57.
  2. Bortner, C.D. et al. (1995) Trends Cell Biol 5, 21-6.
  3. Gavrieli, Y. et al. (1992) J Cell Biol 119, 493-501.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 6
#48513

TUNEL Assay Kit (Fluorescence, 594 nm)

Immunohistochemistry Image 1: TUNEL Assay Kit (Fluorescence, 594 nm) Expand Image
Confocal analysis of paraffin-embedded human colon carcinoma using TUNEL Assay Kit (Fluorescence, 594 nm) (red) followed by staining with Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664 (green) and DAPI #4083 (blue).
Immunohistochemistry Image 2: TUNEL Assay Kit (Fluorescence, 594 nm) Expand Image
Confocal analysis of paraffin-embedded HT-29 xenograft using TUNEL Assay Kit (Fluorescence, 594 nm) (red) and DAPI #4083 (blue).
Immunofluorescence Image 1: TUNEL Assay Kit (Fluorescence, 594 nm) Expand Image
Confocal analysis of HeLa cells, treated with Staurosporine #9953 (1 µM, 3 hr; left, positive) or untreated (right, negative), using TUNEL Assay Kit (Fluorescence, 594 nm) (red) followed by staining with Cleaved Caspase-3 (Asp175) Antibody #9661 (green) and DAPI #4083 (blue).
Immunofluorescence Image 2: TUNEL Assay Kit (Fluorescence, 594 nm) Expand Image
Confocal analysis of a fixed frozen E14.5 mouse digit at low magnification (left) and high magnification (right) using TUNEL Assay Kit (Fluorescence, 594 nm) (red) followed by staining with Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664 (green) and DAPI #4083 (blue).
Flow Cytometry Image 1: TUNEL Assay Kit (Fluorescence, 594 nm) Expand Image
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 3 hr; green), unlabeled (dashed lines) or labeled with TUNEL Assay Kit (Fluorescence, 594 nm) (solid lines).