APO-BRDU-IHC (TUNEL) Apoptosis Kit

• Applications: ICC/IF, IHC

APO-BRDU-IHC (TUNEL) Apoptosis Kit Summary

• Description: The 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 the instructions and reagents required for measuring apoptosis in cells including; 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 anti-BrdU antibody for labeling DNA breaks, horseradish peroxidase streptavidin conjugate, DAB, H2O2/Urea tablets for color generation and methyl green solution for counterstaining the cells.
• Kit Type: Apoptosis Kit

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
• Application Notes: Please centrifuge the TdT Enzyme for 30 seconds, upon warming, to ensure all liquid is at the bottom of the tube. NOTE:- The positive and negative control cells supplied in the kit are from a cytospin preparation and do not need to be deparaffinized.

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Alternate Names for APO-BRDU-IHC (TUNEL) Apoptosis Kit

• APO BRDU
• APO-BRDU
• APO-BRDU-IHC
• TUNEL assay kit

Background

One of the most easily measured features of apoptotic cells is the break-up of the genomic DNA by cellular nucleases. These DNA fragments can be extracted from apoptotic cells and result in the appearance of "DNA laddering" when the DNA is analyzed by agarose gel electrophoresis. The DNA of non-apoptotic cells which remains largely intact does not display this "laddering" on agarose gels during electrophoresis. The large number of DNA fragments appearing in apoptotic cells results in a multitude of 3'-hydroxyl ends in the DNA. This property can be used to identify apoptotic cells by labeling the 3'-hydroxyl ends of the DNA with bromolated deoxyuridine triphosphate nucleotides (Br-dUTP). The enzyme terminal deoxynucleotidyl transferase (TdT) catalyzes this template independent addition of the Br-dUPT's to the 3'-hydroxyl ends of double- or single-stranded DNA with either blunt, recessed or overhanging ends. A substantial number of these sites are available in apoptotic cells providing the basis for the method utilized in the APO-BRDU-IHC (TUNEL) Apoptosis Kit. Non-apoptotic cells do not incorporate significant amounts of the Br-dUTP owing to the lack of exposed 3'-hydroxyl DNA ends. After labeling the 3'-hydroxyl ends of the DNA with Br-dUTP, an anti-BrdU antibody directly conjugated to biotin is attached to the Br-dUTP's. Horseradish peroxidase conjugated to avidin is attached to the biotin labeled Br-dUTP's. Then a standard contrast microscope stain reaction, DAB-H2O2-Urea, is used to label the incorporated Br-dUTP's. Recent evidence has demonstrated that Br-dUTP is more readily incorporated into the genome of apoptotic cells than are the deoxynucleotide triphosphates complexed to larger ligands like fluorescein, biotin or digoxigenin. This greater incorporation gives rise to a stronger flow cytometry signal when the Br-dUTP sites are identified by a fluorescein labeled anti BrdU monoclonal antibody and may increase the signal to noise ratio in this immunohistochemical staining method.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for APO-BRDU-IHC (TUNEL) Kit (NBP2-31164)(11)

Publications using NBP2-31164

• Kervancioglu Demirci E, Onen EA, Sevic Yilmaz E et al. SARS-CoV-2 Causes Brain Damage: Therapeutic Intervention with AZD8797 Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada 2023-11-15 [PMID: 37967299]
• Choudhary R, Kumar M, Katyal A 12/15-Lipoxygenase debilitates mitochondrial health in intermittent hypobaric hypoxia induced neuronal damage: An in vivo study Redox biology 2021-12-30 [PMID: 34979449]
• Lu Q, Zhang Y, Kasetti RB et al. Nervilifordin F alleviates intestinal ischemia/reperfusion-induced acute lung injury via inhibiting inflammasome and mTOR pathway Invest Ophthalmol Vis Sci 2020-10-08 [PMID: 33085740] (Mouse)
• Engelbertsen D, Autio A, Verwilligen RAF et al. Increased lymphocyte activation and atherosclerosis in CD47-deficient mice Sci Rep 2019-07-23 [PMID: 31337788]
• An, Y;Wang, P;Xu, P;Tung, HC;Xie, Y;Kirisci, L;Xu, M;Ren, S;Tian, X;Ma, X;Xie, W; An unexpected role of cholesterol sulfotransferase and its regulation in sensitizing mice to acetaminophen induced liver injury Mol. Pharmacol. 2019-04-03 [PMID: 30944208] (ICC/IF, Human)
• Bu Y, Yoshida A, Chitnis N et al. A PERK-miR-211 axis suppresses circadian regulators and protein synthesis to promote cancer cell survival. Nat. Cell Biol. 2018-01-01 [PMID: 29230015] (ICC/IF)
• Guo G, Yu M, Xiao W et al. Local activation of p53 in the tumor microenvironment overcomes immune suppression and enhances antitumor immunity. Cancer Res. 2017-03-09 [PMID: 28280037]
• Wagh PR Parkin and PINK1 mitigates severity of acute pancreatitis and accelerates recovery of exocrine pancreas Thesis
• Ishiguro S, Upreti D, Robben N et al. Water extract from Euglena gracilis prevents lung carcinoma growth in mice by attenuation of the myeloid-derived cell population Biomed. Pharmacother. 2020-04-28 [PMID: 32361165]
• Ishiguro S, Robben N, Burghart R et al. Cell Wall Membrane Fraction of Chlorella sorokiniana Enhances Host Antitumor Immunity and Inhibits Colon Carcinoma Growth in Mice Integr Cancer Ther 2020-01-01 [PMID: 32009489]
• ZHANG SM, CAI XF, MA YL Effect of rosiglitazone on myocardial injury in septic rats through NF-kappa B pathway European Review for Medical and Pharmacological Sciences 2020-01-01 [PMID: 31957860] (Rat)

FAQs for APO-BRDU-IHC (TUNEL) Kit (NBP2-31164). (Showing 1 - 1 of 1 FAQs).

1. How many tests can it be done?
   • This kit comes with enough material for 50 assays and includes 2 positive & 2 negative control slides.