Product Category
FLARE™ Assay Kits
Fragment Length Analysis using Repair Enzymes
FLARE™ Assay Kits provide the reagents necessary to screen for DNA damage in single cells using specific enzymes that cleave at sites of DNA damage in conjunction with the CometAssay™. To assess the type or extent of DNA damage induced by a putative mutagen, drug, or treatment regimen, treated cells are harvested and immobilized in a layer of low melting point agarose on the FLARE slide. The cells are gently lysed and subsequently incubated with a kit-specific enzyme which recognizes a particular type of DNA damage (as outlined below) and cleaves the DNA. After treatment with alkali to denature the DNA, the samples are submitted to neutral gel electrophoresis, and stained using SYBR® Green I. Denatured, cleaved DNA fragments migrate out of the cell during electrophoresis, while undamaged, supercoiled DNA remains within the confines of the nuclear cell membrane. The extent of the DNA damage can be assessed qualitatively, or quantitatively using image analysis software to calculate tail length and tail moment.
All FLARE Assay kits contain:
- 25 FLARE Slides
- 25X FLARE Buffer 2
- REC™ Dilution Buffer
- 100X BSA Additive
- 100X Cation Solution
- Lysis Solution
- Comet LMAgarose
- SYBR Green Concentrate
- 75 Tests
In addition, each kit contains a specific cleavage enzyme:
- Human OGG1 (Catalog # 4130-100-FK):
Catalyzes the removal of 8-oxoguanine and DNA-containing formamidopyrimidine moieties.
- Chorella virus PDG (Catalog # 4065-100-FK):
Recognizes cis-syn and trans-syn cyclobutane pyrimidine dimers induced by UV irradiation. Also cleaves AP sites on double-, and single-stranded DNA.
- Fpg (Catalog # 4040-100-FK):
Catalyzes the excision of multiple forms of damage bases, including 8-oxoguanine, 5-hydroxycytosine, 5-hydroxyuracil, aflatoxin-bound imidazole ring-opened guanine, imidazole ring-opened N-2-aminofluorene-C8-guanine, and open ring forms of 7-methylguanine.
- T4-Endonuclease V (Catalog # 4055-100-FK):
Cleaves the cis-syn isomer of cyclobutane pyrimidine dimers induced by UV irradiation, and AP sites.
- UVDE (Catalog # 4100-100-FK):
Repairs bipyrimidine DNA photoproducts, such as cyclobutane pyrimidine dimers and photoproducts, and cleaves immediately 5’ to the photoproduct site.
Additional FLARE Slides are also available in packs of 25 (Catalog # 3950-075-02).
FLARE and CometAssay are trademarks of Trevigen, Inc.