APC BrdU Flow Kit
- Brand BD Pharmingen™
- Regulatory Status RUO
Regulatory Status Legend
The immunofluorescent staining of incorporated bromodeoxyuridine (BrdU) and flow cytometric analysis provide a high-resolution technique to determine the frequency and nature of individual cells that have synthesized DNA. In this method, BrdU (an analog of the DNA precursor thymidine) is incorporated into newly synthesized DNA by cells entering and progressing through the S (DNA synthesis) phase of the cell cycle.The incorporated BrdU is stained with specific anti-BrdU fluorescent antibodies. The levels of cell-associated BrdU are then measured by flow cytometry. Often, staining with a dye that binds to total DNA such as 7-amino-actinomycin D (7-AAD) is coupled with immunofluorescent BrdU staining. With this combination, two-color flow cytometric analysis permits the enumeration and characterization of cells that are actively synthesizing DNA (BrdU incorporation) in terms of their cell cycle position (ie, G0/1, S, or G2/M phase defined by 7-AAD staining intensities).
|Fluorochrome-conjugated anti-BrdU Antibody||One 65-µL vial|
|BD Cytofix/Cytoperm™ Buffer||One 25-mL bottle|
|BD Perm/Wash™ Buffer (10X)||Two 25-mL bottles|
|BD Cytoperm™ Permeabilization Buffer Plus||One 10-mL bottle|
|7-AAD||One 1-mL vial|
|BrdU (10 mg/mL)||Five 0.5-mL vials|
|DNase||Five 300-µL vials|
Preparation and Storage
Antibody, buffers, and 7-AAD.
Store the fluorochrome-conjugated anti-BrdU antibody, BD Cytofix/Cytoperm Buffer, BD Perm/Wash Buffer, BD Cytoperm Permeabilization Buffer Plus, and 7-AAD at 2 to 8°C. Keep the fluorochrome-conjugated anti-BrdU antibody and 7-AAD protected from light.
Unused portions of 1X BD Cytofix/Cytoperm Buffer can be stored at 2 to 8°C.
BrdU and DNase.
Store the BrdU and DNase at –80°C.
The BrdU solution has been shown to be stable for up to 4 months at 2 to 8°C, or it can be refrozen. Avoid multiple freeze-thaw cycles.
DNase stock solution (1 mg/mL) may be refrozen once before it loses activity.