11299964001
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
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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.
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).
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 Statements
Precautionary Statements
Hazard Classifications
Acute Tox. 4 - Acute Tox. 4 Dermal - Eye Irrit. 2 - Flam. Liq. 3 Inhalation - 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
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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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