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  • : APT116
  • : 50 assays
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Trade Name:
Chemicon (Millipore)
50 assays
Product Overview:
The 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.
The 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:
Species Reactivity:
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.
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Storage Conditions:
Precautions 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.
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Materials Required but Not Delivered:
1. 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
  • 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 Methods: