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ApoBrdU DNA Fragmentation Assay Kit

based on 16 citations in multiple journalsApoBrdU DNA Fragmentation Assay Kit165 5
Detects DNA Fragmentation by FACS or FL
Catalog #: K401

In stock

$615.00

Product Details

Cat # +Size K401-60
Size 60 assays
Detection Method Flow cytometry (Ex/Em = 488/520 nm for FITC, and 488/623 nm for PI) and fluorescence microscopy (FITC and rhodamine filters)
Species Reactivity Mammalian
Applications 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.
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-FITC Antibody
• PI/RNase Staining Buffer
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Internucleosomal DNA fragmentation is a hallmark of apoptosis in mammalian cells. BioVision’s Apo-BrdU 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 fluorescein labeled anti-BrdU monoclonal antibody.


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We recently purchased your Apo-BrdU DNA Fragmentation Assay Kit (catalogue #K401-60) for use on our rat brain tissue sections and we are having issues with it. While we are able to detect the propidium iodide labeling without problem, we have not been able to label any TUNEL. Even when using DNase as a positive control and when leaving off the DNA labeling solution step as a negative control, all of our sections look alike. I have tried many different things and I see no reason why it shouldn't be working. Any suggestion?
There are a couple of things that you can do to improve. 1. The perfusion method may not have been sufficient to fix the tissues. Improper fixing might result in loss of DNA that are not chemically fixed in place. Try fixing the tissues post cryosection2. Pre-wetting the tissue can help before adding the labeling mix. Incubate the tissue with just 1X Reaction buffer diluted in ddH20. 3. Increasing the incubation time >1.5hr can also help in getting better staining. 4. do a DNase treatment with 1µg/ml for 30 min at 37°C. 4. Try diluting PI solution 1:5 in PBS.
How should we process the positve and negative controls with our tissue sections and there is nothing in the instructions about the controls. Could you please clarify?
The positive and negative controls are included just to ensure that the kit components are working fine and also adjust the parameters/filter set up for fluorescence microscopy. You can process these cells in the same way you process sample cells . Please take a lot at Section B. Detection by Flow Cytometry and Fluorescence Microscopy: The procedures can be used for both control cells and your testing cells.
From which species the anti-BrdU FITH antibody from kit K401 is?
This antibody is from mouse.
Can the kit detect detect cyclobutane pyrimidine?
ApoBrdU DNA fragmentation kit helps measure the extent of DNA fragmentation, not specific modified DNA-derived nucleotides like cyclobutane pyrimidine.
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.
Kuo, Jinn-Rung et al. (2016) Cisplatin-induced regulation of signal transduction pathways and transcription factors in p53-mutated subclone variants of hepatoma cells: Potential application for therapeutic targeting, Oncol Lett. 2016 Nov;12(5):3723-3730.
Malik, Salma et al. (2016) Therapeutic Potential and Molecular Mechanisms of Emblica officinalis Gaertn in Countering Nephrotoxicity in Rats Induced by the Chemotherapeutic Agent Cisplatin, Front Pharmacol. 2016 Oct 3;7:350.
Shang, Hung-Sheng et al. (2016) Curcumin causes DNA damage and affects associated protein expression in HeLa human cervical cancer cells, Oncol Rep. 2016 Oct;36(4):2207-15.
Bharti, Brij et al. (2016) Level of PAX5 in differential diagnosis of non-Hodgkin's lymphoma, Indian J Med Res. 2016 May;143(Supplement):S23-S31.
Sun et al., Change of Recipient Corneal Endothelial Cells After Non-Descemet's Stripping Automated Endothelial Keratoplasty in a Rabbit Model. Invest. Ophthalmol. Vis. Sci., Dec 2014; 55: 8467 - 8474.
For more citations of this product click here