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ApoBrdU-IHC DNA Fragmentation Assay Kit

based on 17 citations in multiple journalsApoBrdU-IHC DNA Fragmentation Assay Kit175 5
Two-Color TUNEL Assay
Catalog #: K403

In stock

$615.00

Product Details

Cat # +Size K403-50
Size 50 assays
Species Reactivity Mammalian
Applications The TUNEL-based immunohistochemical staining kit provides complete reagents including positive and negative control slides 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 • Control Slides-pos/neg
• Blocking Buffer
• H₂O₂/Urea Tablets
• Protease K
• DAB Tablets
• TdT Enzymes
• Br-dUTP
• 200X Conjugate
• 5X Reaction Buffer
• Anti-BrdU-Biotin Antibody
• Methyl Green
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-IHC™ Kit is a two-color TUNEL (Terminal deoxynucleotide transferase dUTP Nick End Labeling) assay for labeling DNA breaks to detect apoptotic cells by immunohistochemistry. The kit contains positive/negative control slides for assessing reagent performance; reaction and blocking buffers for processing individual steps in the assay; proteinase K; terminal deoxynucleotidyl tranferase enzyme (TdT), bromodeoxyuridine triphosphate (Br-dUTP), biotin labeled antiBrdU antibody for labeling DNA breaks, horseradish peroxidase streptavidin conjugate, DAB, H₂O₂/Urea tablets for color generation and methyl green solution for counter staining the cells.


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All my cells both treated and control is showing positive PI stain. Why so?
The reason why PI is staining all sample types is because the Reaction buffer and the Rinse buffer have a detergent in them, Triton-X. You need the detergent to allow the probe to enter the nucleus to stain the DNA. So if you use the kit, all cells should stain positive for PI, and PI is just being used as a counterstain.
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.
In the last step of the protocol (step 10), there is no description how the cells are transferred to slides for fluorescence microscopy. May I know how the cells are transferred to slides?
We take 50-100ul of cells and put it on the slide, invert a coverslip on it and seal with nail-polish. Poly-lysine treated slides are better.
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.
Nunez-Duran E et al., (2017) Protein kinase STK25 aggravates the severity of non-alcoholic fatty pancreas disease in mice, J Endocrinol, 2017, 234: 15-27
Chunlai F et al., (2017) Dexmedetomidine attenuates lipopolysaccharide-induced acute lung injury by inhibiting oxidative stress, mitochondrial dysfunction and apoptosis in rats, Mol Med Reports, 2017, 15(1): 131-138
Kim, Jin Young et al. (2017) Bilirubin nanoparticle preconditioning protects against hepatic ischemia-reperfusion injury, Biomaterials. 2017 Jul;133:1-10.
Peng, Y.-B. et al. (2017) The study of the relationship between aberrant expression of hot shock protein 70 (HSP70) and spontaneous abortion, Eur Rev Med Pharmacol Sci. 2017 Feb;21(4):652-656
Suchal, Kapil et al. (2017) Protective effect of mangiferin on myocardial ischemia-reperfusion injury in streptozotocin-induced diabetic rats: role of AGE-RAGE/ MAPK pathways, Sci Rep. 2017 Feb 9;7:42027.
For more citations of this product click here
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