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Roche

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

sufficient for ≤100 tests, storage temp.:-20°C

Synonym(s):

5-BrdU, 5-Bromo-2-deoxyuridine

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About This Item

UNSPSC Code:
41116133

Quality Level

usage

sufficient for ≤100 tests

manufacturer/tradename

Roche

storage temp.

−20°C

General description

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.
Immunohistocytochemical assay for the detection of 5-bromo-2′-deoxy-uridine (BrdU) incorporated into cellular DNA.

Specificity

Anti-BrdU monoclonal antibody specifically binds to 5-bromo-2′-deoxy-uridine, and shows cross-reactivity with 5-iodo-2′-deoxy-uridine (10%). Anti-BrdU shows no cross-reactivity with 5-fluoro-2′-deoxy-uridine or any endogenous cellular component, such as thymidine or uridine.

Application

5-Bromo-2′-deoxy-uridine Labeling and Detection Kit II has been used in:
  • labeling of tooth roots for histology
  • immunostaining of mice frontal sections
  • immunofluorescence imaging of hepatocellular carcinoma sections

The kit is used for the detection of BrdU incorporated into cellular DNA by immunohistocytochemistry.

Features and 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.

Packaging

1 kit containing 7 components.

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.

Preparation Note

Working concentration: Working concentration of the labeling reagent corresponds to the WC of the In Situ Cell Proliferation Kits.
Sample material:
Cell culture:
adherent cells, suspension cells, organ or explant cultures. Frozen or paraffin-embedded tissue sections (after in vivo labeling).

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

Product No.
Description

  • BrdU Labeling Reagent 1,000x concentrated

  • Washing Buffer concentrate 10x concentrated

  • Incubation Buffer

  • Anti-BrdU antibody, contains nucleases for DNA denaturation

  • Anti-mouse Ig-alkaline Phosphatase antibody

  • NBT

  • BCIP

Signal Word

Danger

Hazard Classifications

Acute Tox. 4 - Acute Tox. 4 Inhalation - Eye Irrit. 2 - Flam. Liq. 3 Dermal - Muta. 1B - Repr. 1B - Skin Sens. 1

Storage Class Code

3 - Flammable liquids

WGK

WGK 2

Flash Point(F)

136.4 °F

Flash Point(C)

58 °C


Certificates of Analysis (COA)

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Articles

Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.

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