How do you do a BrdU assay?

How do you do a BrdU assay?

Label cells with BrdU

1. Culture cells in appropriate vessel for microscopy.
2. Remove culture medium from cells and replace with BrdU labeling solution.
3. Incubate cells at 37°C for 2 hours.
4. Remove labeling solution and wash two times with PBS.
5. Wash with PBS (3 times, 2 minutes each)

What is BrdU assay?

Bromodeoxyuridine (BrdU) incorporation assays have long been used to detect DNA synthesis in vivo and in vitro. The key principle of this method is that BrdU incorporated as a thymidine analog into nuclear DNA represents a label that can be tracked using antibody probes.

How is BrdU tested?

Detect incorporated BrdU

1. Remove this solution and add 1 mL of antibody staining buffer.
2. Add anti-BrdU primary antibody.
3. Incubate overnight at room temperature.
4. Wash with Triton X-100 permeabilization buffer (3 times, 2 minutes each)
5. Add fluorescently labeled secondary antibody.
6. Incubate one hour at room temperature.

What is the difference between EdU and BrdU?

EdU assay. Unlike assays using BrdU staining, Click-iT EdU assays are not antibody-based and therefore do not require DNA denaturation for detection of the incorporated nucleoside.

What kind of cells does BrdU stain for?

BrdU (Bromodeoxyuridine / 5-bromo-2′-deoxyuridine) is an analog of the nucleoside thymidine used in the BrdU assay to identify proliferating cells. BrdU labeling can be performed in vitro for cell lines and primary cell cultures, or in vivo for labeling cells within a living animal.

What is BrdU a marker of?

BrdU is not a marker of the S-phase of the cell cycle. As a thymidine analog, it is a marker of DNA synthesis.

What is EdU assay?

EdU Assay / EdU Staining Proliferation Kit (iFluor 488) ab219801 provides a sensitive and robust method to detect and quantify cell proliferation in live mammalian cells using flow cytometry or fluorescence microscopy.

What is the purpose of BrdU in the study of cell cycle?

BrdU is commonly used to study cell proliferation in living tissues and has been studied as a radiosensitizer and diagnostic tool in people with cancer….Bromodeoxyuridine.

What kind of cells does BrdU identify?

What are BrdU cells?

Bromodeoxyuridine (BrdU) is a thymidine analog that incorporates DNA of dividing cells during the S-phase of the cell cycle. As such, BrdU is used for birth dating and monitoring cell proliferation.