APT116 | Apo-BRDU IHC Kit

APT116
50 assays  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsDetection Methods
      IHCChromogenic
      Description
      Catalogue NumberAPT116
      Brand Family Chemicon®
      Trade Name
      • Apo-BrdU
      • Chemicon
      DescriptionApo-BRDU IHC Kit
      OverviewThe CHEMICON APO-BRDU-IHC™ Kit is a two color staining method for labeling DNA breaks and total cellular DNA to detect apoptotic cells by immunohistochemistry (Li et al. 1995). The kit contains the instructions and reagents required for measuring apoptosis in cells including: positive and negative control slides for assessing reagent performance; reaction and blocking buffers for processing individual steps in the assay; terminal deoxynucleotidyl tranferase enzyme (TdT), bromodeoxyuridine triphosphate (Br-dUTP), biotin-labeled anti-BrdU antibody for labeling DNA strand breaks, HRP-streptavidin conjugate, DAN, H2O2/Urea tablets for color generation, and methyl green solution for counter staining the cells.
      Materials Required but Not Delivered1. Xylene - 100%

      2. Ethanol - 100%, 90%, 80%, 70% v/v with dH2O

      3. Methanol - 100%

      4. Hydrogen Peroxide - 30%

      5. PBS or Tris-buffered saline (1X TBS, 20 mM Tris pH 7.6, 140 mM NaCl)

      6. 10 mM Tris, pH 8.0

      7. 1 mM MgSO4 in 1X TBS (for use in generating positive control)

      8. DNase I (for use in generating positive control)

      9. 1-4% Formaldehyde in PBS, pH 7.4

      10. Distilled Water

      11. Coplin jars with slide holders

      12. Beaker for rinsing slides

      13. Slide humidifying chamber

      14. Glass cover slips

      15. Mounting media

      16. Microscope

      17. Micro-pipettor with disposable tips

      18. Microcentrifuge tubes

      19. Parafilm® (Registered trademark of American National Can Company)

      20. Absorbent wipes

      21. Crushed ice
      References
      Product Information
      Components
      • Positive/Negative Control Slides, Natural box
      • Note: 2 slides, each containing a mixture positive and negative control cells.
      • Blocking Buffer, White cap
      • H2O2/Urea Tablets, Amber vial
      • Proteinase K, Pink cap
      • DAB Tablets, Amber vial
      • TdT Enzyme, Yellow cap
      • Br-Dutp, Violet cap
      • 200X Streptavidin-HRP Conjugate, Black cap
      • 5X Reaction Buffer, Green cap
      • Biotin-labeled BrdU mAb, Orange cap
      • Methyl Green solution, Natural cap,
      • The APO-BRDU-IHC™ Kit is shipped in one container and should be stored at -20°C upon arrival.
      • Note: After the kit is defrosted before first use, both the 5X Reaction Buffer and the Biotin-labeled anti-BrdU antibody should be stored at 2-8°C. Also, the Methyl Green solution should be stored at room temperature. DO NOT RE-FREEZE any of these components once thawed.
      • Reagent bottles have color coded caps to aid in their identification. Sufficient reagents are provided to process 50 slides including 2 positive/negative control slides. The control cells are derived from a human promyelocitic leukemia cell line (HL60) and incubated with 0.5 mg/mL actinomycin D for 19 hours to induce apoptosis.
      Detection methodChromogenic
      Applications
      ApplicationThe APO-BRDU-IHC Kit is a two color staining method for labeling DNA breaks & total cellular DNA to detect apoptotic cells by IHC.
      Key Applications
      • Immunohistochemistry
      Biological Information
      Species Reactivity
      • All
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsPrecautions and Warnings:

      1. The components of this kit are for Research Use only and are not intended for diagnostic procedures.

      2. After initial defrosting, store the 5X Reaction Buffer and Biotin-labeled anti-BrdU antibody at 2-8ºC, and the Methyl Green solution at room temperature. DO NOT RE-FREEZE any of these components once thawed.

      3. TdT Enzyme will not freeze at -20oC, because it is in 50% (v/v) glycerol solution.

      4. The 5X Reaction buffer and Br-dUTP solution contain cacodylic acid as a buffer. The Biotin-labeled antibody contains 0.05% sodium azide as a preservative. The DAB tablets contain diaminobenzidine. These materials are harmful if swallowed. Avoid skin contact with any of these components; wash immediately with water.

      5. Upon warming the components, centrifuge the tubes for 30 seconds to force all the liquid to the bottom of the tube.

      6. Gloves, lab coat and protective eyewear should be worn while staining.
      Packaging Information
      Material Size50 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      References

      Reference overviewPub Med ID
      Labelling DNA strand breaks with BrdUTP. Detection of apoptosis and cell proliferation.
      Li, X and Darzynkiewicz, Z
      Cell Prolif., 28: 571-9 (1995)  1995

      Show Abstract
      8555370 8555370

      Instructions for Use

      Title
      APO-BRDU-IHC¿ Kit for Measuring Apoptosis by Dual Color Immunohistochemistry