ApoBrdU Red DNA Fragmentation Kit

Detects DNA Fragmentation by FACS or FL

WARNING: This product can expose you to chemical including Cacodylic acid, which is known to the State of California to cause cancer. For more information go to
Catalog #: K404 | abID: ab66110

Product Details

abID ab66110
Cat # +Size K404-60
Size 60 assays
Detection Method Flow cytometry (Ex/Em = 488/576 nm for BrdU-Red and 488/655 nm for 7-AAD).
Species Reactivity Mammalian
Applications The TUNEL-based immunohistochemical staining kit provides complete reagents including positive and negative control cells for convenient detection of DNA fragmentation
Features & Benefits • Simple procedure; takes ~ 3 hours
• Fast and convenient
• The assay is sensitive and stable
• The TUNEL-based assay kit provides complete components including positive and negative control cells for convenient detection of DNA fragmentation in cultured cells and tissue sections.
Kit Components • Positive Control Cells
• Negative Control Cells
• Wash Buffer
• Reaction Buffer
• TdT Enzymes
• Br-dUTP
• Rinse Buffer
• Anti-BrdU-Red Antibody
• PI/RNase Staining Buffer
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells. BioVision’s Apo-BrdU-Red In Situ DNA Fragmentation Assay Kit provides complete components including positive and negative control cells for conveniently detecting DNA fragmentation by fluorescence microscopy or flow cytometry. The kit utilizes Br-dUTP (bromolated deoxyuridine triphosphate nucleotides) which is more readily incorporated into DNA strand breaks than other larger ligands (e.g., fluorescein, biotin or digoxigenin). The greater incorporation gives rise to brighter signal when the Br-dUTP sites are identified by a Red fluorescence labeled anti-BrdU monoclonal antibody. The assay is suitable for studying apoptosis with GFP transfected cells.

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We used K404 according to the protocol and did not observe any signal under the recommended settings (Ex/Em = 488/576). However, we were able to observe signal after changing the excitation to 555 nm (emission remained the same). Do you have any insights as to why this may be happening, and would you expect the signal they observed to be correct for this kit?
Actually, this is absolutely correct (except that they couldn’t excite at 488 is a little troubling) because the Red- anti-BrdU actually has “optimal” excitation at 544 nm. However, we have found that it still excites at 488 nm wavelength, which allows it to use it on a typical flow cytometer and especially on a single laser system such as the BD FACScan, for which it was originally developed. The emission should not change as the customer discovered. The same is true for the counter-stain used in the kit. It excites very well at 488 (standard flow cytometers) but its optimal excitation is at 546 nm and emission at 647 nm.
What is the spectrum of excitation and emission of the BrdU-Red?
The BrdU-Red monoclonal antibody emits fluorescence at 576nm when excited at 488nm. The wavelengths can shift +/-10 to 20nm based on the instrument.
Can this assay differentiate between necrosis/apoptosis?
TUNEL assay is positive for both necrotic and apoptotic cells and hence cannot distinguish. You can run DNA gel to differentiate between them- while necrotic cells will show a smear due to non-specific cleavage, apoptotic cells has activated endonucleases that cleaves the DNA into fragments of approximately 180-200bp that looks like a ladder on the DNA gel.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Would you please provide any protocol regarding how to adhere the fixed cells (Positive Control provided in the kit) or unfixed cells on the slide?
For fixed cells, as far as we know, you have to use cytospin to put them on the slide for microscopy. Otherwise, you may go with flow cytometry as the detection method.

For non-fixed cells, you can use poly-lysine coated slides/coverslips. Poly-lysine will not work with fixed cells as it based on the interaction between positively charged poly-lysine and negatively charged cells or proteins and in fixed cells, this interaction does not work. Please use a standard protocol for coating the slides/coverslips with poly-lysine.
Sara Della Torre, et al., (2018) Short-Term Fasting Reveals Amino Acid Metabolism as a Major Sex-Discriminating Factor in the Liver, Cell Metabolism, Aug. 2018, 29909969
Persson, Andrea et al. (2016) Xyloside-primed Chondroitin Sulfate/Dermatan Sulfate from Breast Carcinoma Cells with a Defined Disaccharide Composition Has Cytotoxic Effects in Vitro J Biol Chem. 2016 Jul 8;291(28):14871-82.
Walsh et al., LKB1 inhibition of NF-Description: {kappa}B in B cells prevents T follicular helper cell differentiation and germinal center formation. EMBO Rep., Apr 2015; 10.15252/embr.201439505.
Che et al., Regulatory T Cells Resist Virus Infection-Induced Apoptosis. J. Virol., Feb 2015; 89: 2112 - 2120.
Cassano, M. et. al. Alpha sarcoglycan is required for FGF-dependent myogenic progenitor cell proliferation in vitro and in vivo. Development, Oct 2011; 138: 4523 - 4533.
For more citations of this product click here