BD Cell Cycle Kit
BD Biosciences introduces the BD™ Cell Cycle Kit, our latest addition to the BD Bioimaging Certified Reagents product line. This cost-effective kit lets you simultaneously determine the percentage of cells in M and S phase using a one-step staining reaction. Further multiplexing can be accomplished by using additional probes for a true high-content imaging assay.
The BD Cell Cycle Kit uses antibodies that are directly conjugated with fluorochromes emitting in the green (Alexa Fluor® 488) and red (Alexa Fluor® 647) channels. This allows for an additional BD Bioimaging certified reagent in the orange channel (for example, Alexa Fluor® 555) to be multiplexed with the kit antibodies. The kit also uses Hoechst nuclear dye.
Features
- A three-color application using fluorescent antibodies in combination to determine the M and S phases of the cell cycle while allowing you to drop in an additional specificity
- Qualified for use in high content screening (HCS) assays for generating reliable and reproducible results
- A simple protocol requiring no optimization or assay development, thus saving you time
- Validated on the BD Pathway™ 435 and 855 high-content bioimagers, and compatible with other HCS platforms and fluorescence microscopes
| Description | Cat. No. |
|---|---|
| BD Cell Cycle Kit (96-well plate or 384-well plate) | 558662 |
Figure 1. U-2 OS cells were either left untreated, or were treated with aphidicolin (250 ng/mL) for one hour or with colchicine (500 ng/mL) for 16 hours prior to BrdU loading.
Aphidicolin, a DNA polymerase inhibitor, blocks cells in early S phase, while colchicine, a microtubule polymerization inhibitor, blocks cells in M phase. Cells were stained according to the BD Cell Cycle Kit protocol. In addition to the two antibodies provided in the kit, an Alexa Fluor® 555 mouse anti-β-tubulin antibody (Cat. No. 558608) was used at the same time to monitor the microtubule depolymerization. Representative 20x images are shown. Hoechst staining is pseudocolored blue, BrdU staining is pseudocolored red (appears pink when co-localized with blue), Histone H3 (pS28) staining is pseudocolored yellow (appears white when co-localized with blue), and β-tubulin staining is pseudocolored green. Image analysis (data not shown) revealed that untreated U-2 OS cells (Panel A) had approximately 30% cells in S phase (pink) and approximately 3% cells in M phase (yellow/white). Aphidicolin treatment (Panel B) blocked cells from entering S phase, but there was no effect on β-tubulin or on cells entering M phase. Conversely, no β-tubulin staining (cytoplasmic intensity was measured using an 8-pixel ring dilated out from the nuclear mask identified by the Hoechst stain) was detected when cells were treated with colchicine (Panel C) and the percentage of cells in M phase increased to 50%.