Product Catalog

5-Bromo-2'-deoxy-uridine Labeling and Detection Kit II

Immunocyto/histochemical assay for the detection of 5-bromo-2'-deoxy-uridine (BrdU) incorporated into cellular DNA

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112999640011 kit (100 tests)Order and Price Information
 


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Application

The kit is used for the detection of BrdU incorporated into cellular DNA by immunocyto/histochemistry.

Benefits

  • Safe: No radioisotopes are used
  • Easy to perform: Follows a standard immunohistochemistry protocol
  • Sensitive: Denaturation of DNA with nucleases allows for highly sensitive detection of BrdU
  • Flexible: Allows double-labeling protocols

Product Description

Sample material: Cell culture: adherent cells, suspension cells, organ or explant cultures. Frozen or paraffin-embedded tissue sections (after in vivo labeling).
Specificity: Anti-BrdU monoclonal antibody specifically binds to 5-bromo-2'-deoxy-uridine, and shows crossreactivity with 5-iodo-2'-deoxy-uridine (10%). Anti-BrdU shows no crossreactivity with 5'-fluoro-2'-deoxy-uridine or any endogenous cellular component, such as thymidine or uridine.

Background Information

Cell proliferation may be studied by monitoring the incorporation of a radioisotope, [3H]-thymidine, into cellular DNA, followed by autoradiography.
Alternatively, 5-bromo-2'-deoxy-uridine (BrdU) may be used instead of thymidine. Cells that have incorporated BrdU into DNA are easily detected using a monoclonal antibody against BrdU and an enzyme- or fluorochrome-conjugated second antibody.

Contents

  1. BrdU Labeling Reagent, 1,000x conc.,  sterile
  2. Washing Buffer concentrate, 10x
  3. Incubation Buffer
  4. Anti-BrdU, containing nucleases for DNA denaturation
  5. Anti-mouse Ig-alkaline Phosphatase
  6. NBT
  7. BCIP

Principle

Samples prelabeled with BrdU are fixed with ethanol, then incubated with a monoclonal antibody to BrdU, which contains an optimized mixture of nucleases. These nucleases generate single-stranded DNA fragments that allow binding of the antibody to BrdU. Next, an alkaline phosphatase (AP)-labeled antibody to mouse immunoglobulin is added, then bound to the anti-BrdU antibody. The sample is then incubated with the AP substrate and NBT/ BCIP, which is metabolized to form a colored reaction product. The sample is evaluated using a phase-contrast microscope.

Figure 1: Principle of the BrdU Labeling and Detection Kit II (AP).

Additional Information


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