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: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:
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
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Western blot analysis of whole cell lysates probed with Goat Anti-NPM Antibody (VPA00058) followed by detection with HRP conjugated Donkey Anti-Sheep/Goat IgG (1/10,000, STAR88P) and visualized on the ChemiDoc MP with 17 second exposure. Arrow points to NPM (molecular weight 38 kDa).
Nucleophosmin, abbreviated as NPM, belongs to the nucleophosmin/nucleoplasmin family of chaperones. NPM is ubiquitously expressed in mammalian cells and shuttles between the nucleus and cytoplasm to regulate cellular processes, such as transport of pre-ribosomal particles, ribosome biogenesis, genome stability, regulation of transcription factors including p53 activity and the “master regulator” of cell cycle gene expression Forkhead box protein M1 (FOXM1). Interaction of NPM with FOXM1 modulates both the level and localization of FOXM1. Overexpression of NPM results in increased cell growth and proliferation.
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Western blot analysis of whole cell lysates probed with Mouse Anti-ROCK1 Antibody (VMA00614) followed by detection with HRP conjugated Goat Anti-Mouse IgG (1/10,000, STAR207P) and visualized on the ChemiDoc MP with 11 second exposure. Arrow points to ROCK1 (molecular weight 185 kDa).
ROCK1 is a serine⁄threonine kinase that acts as a downstream effector of Rho. It is a key regulator of the actin cytoskeleton, cell polarity and cancer cell invasion. Activation of ROCK has been implicated in tumor formation in mice due to its importance in the maintenance of actomyosin contractility and cell proliferation regulation. Strong evidence also suggests a role for ROCK in glucose metabolism. Major downstream ROCK1 substrates include DAPK3, FHOD1, GFAP, JIP3, LIMK1, LIMK2, MLC, PFN1, PPP1R12A. ROCK-1 is a direct cleavage substrate of activated caspase-3.
Applications: Western Blotting
Related Products: Antibodies for kinase research