How do you do a BrdU assay?
Detect incorporated BrdU
- Remove this solution and add 1 mL of antibody staining buffer.
- Add anti-BrdU primary antibody.
- Incubate overnight at room temperature.
- Wash with Triton X-100 permeabilization buffer (3 times, 2 minutes each)
- Add fluorescently labeled secondary antibody.
- Incubate one hour at room temperature.
What is BrdU and how does iT detect mitosis?
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.
What is BrdU incorporation?
Abstract. 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.
What is BrdU and how does it detect mitosis?
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. 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 BrdU a marker of?
As a thymidine analog, it is a marker of DNA synthesis. Therefore, studying neurogenesis with BrdU requires distinguishing cell proliferation and neurogenesis from other events involving DNA synthesis, like DNA repair, abortive cell cycle reentry and gene duplication.
What is high Ki-67?
A result of less than 6% is considered low, 6-10% intermediate, and more than 10% is considered high. Ki-67: Ki-67 is a protein in cells that increases as they prepare to divide into new cells. A staining process can measure the percentage of tumor cells that are positive for Ki-67.
