11299964001

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
• UNSPSC Code: 41116133

Properties

• Quality Level: 100
• usage: sufficient for ≤100 tests
• manufacturer/tradename: Roche
• storage temp.: −20°C

Description

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

• 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

Safety Information

• Pictograms: GHS02,GHS07,GHS08
• Signal Word: Danger
• Hazard Statements: H226,H312 + H332,H317,H319,H340,H360D
• Precautionary Statements: P201 - P210 - P261 - P280 - P308 + P313 - P370 + P378
• 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