Schematic representation of the cell cycle. Ki-67 is expressed during G1, S, G2 and M phases. PCNA is present from G1 through to G2. MCM 2 is critical for replication in G1 while the BrdU method recognizes replicating cells during S phase.
Cell proliferation is the rapid expansion of a cell population due to cell growth and division. This cellular process has to be carefully regulated as the smallest alterations can result in an uncontrolled increase of cell numbers, and eventually cancer. To sustain homeostasis and ultimately prevent carcinogenesis, uncontrolled cell proliferation is often counteracted by an increase in apoptosis.
In eukaryotes the cell cycle can be grouped into three major stages:
Cell cycle deregulation and impaired cell cycle checkpoints are two of the most common causes of aberrant cell proliferation. Cell proliferation can be measured by quantifying protein levels of key proliferation markers such as Ki-67, proliferating cell nuclear antigen (PCNA) and minichromosome maintenance 2 (MCM 2), and also by measuring incorporation of Bromodeoxyuridine (BrdU).
BrdU is a thymidine analog, which is incorporated into the newly synthesized DNA of proliferating cells instead of thymidine when added to cell culture media. The incorporated BrdU can be detected and measured with the help of BrdU specific antibodies.
Learn more about cell proliferation assessments and the key cell proliferation biomarkers Ki-67, MCM2 and PCNA in our Mini-review titled "Understanding the Eukaryotic Cell Cycle - a Biological and Experimental Overview".
alamarBlue® provides a rapid and sensitive way to measure cell proliferation. Just add and measure.
alamarBlue® assays are a convenient and quick way to measure cell proliferation and cytotoxicity in human and animal cell lines, bacteria and fungi. Key advantages over traditional MTT and XTT assays include:
Access our alamarBlue® page for FAQs, online calculators, example calculations, references, protocols or download the alamarBlue® brochure.
Immunostaining of BrdU labeled HeLa cells with Mouse anti-BrdU antibody, clone Bu20a (MCA2483, red). Cytoplasm was stained with Rabbit anti-GAPDH antibody (AHP1628, green). PureBlu™ DAPI (1351303, blue) was used as a nuclear counterstain.
Mouse anti-BrdU antibody, clone Bu20a is one of the most commonly used anti-BrdU antibodies. The product is supported by comprehensive protocols for use in flow cytometry and immunocytochemistry, and 10 publications.
Applications: Flow Cytometry, Immunocytochemistry, Immunofluorescence
Related Products: APO-BrdU™ Flow Cytometry Kit (APO001),
APO-BrdU IHC™ Immunohistochemistry Kit (APO002)
Find out more about BrdU
Staining of MCF-7 breast cancer cells with the HuCAL anti Ki-67 bivalent Fab antibody (HCA006, green). Nuclei were counterstained with Hoechst (blue) and F-actin with Phalloidin (red).
The anti-Ki-67 antibody (HCA006) is a recombinant bivalent Fab antibody selected from the HuCAL GOLD® phage display library for its specificity against the GFKELFQ epitope, which is present eight times in the human Ki-67 protein. For easy detection the Fab fragment is tagged with myc and His protein tags. Due to the absence of an Fc region, HCA006 is highly recommended for the detection of Ki-67 in cells and tissues that are rich in Fc receptors.
Applications: ELISA, Immunofluorescence, Immunohistochemistry, Western Blotting
Related Products: Mouse anti-Penta Histidine Tag:HRP antibody (MCA5995P) and Goat anti Human IgG F(ab’)2:HRP (0500-0099)
Learn more about the HuCAL® technology and antibodies
alamarBlue® is manufactured for Bio-Rad by Trek Diagnostic System. US. patent 5,501,959.
Alexa Fluor® is a registered trademark of Molecular Probes Inc, OR, USA.
Apo-BrdUTM is a registered trademark of Phoenix Flow Systems USA.
HuCAL® and HuCAL® GOLD are registered trademarks of MorphoSys AG.
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