Cell Cycle Chromobody® plasmid (RFP)

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Target Name (Antigen)
  • 53/13
  • CG9193
  • DmPCNA
  • Dmel\\CG9193
  • MUS209
  • Mus209
  • PCNA
  • PCNA/mus209
  • Pcna
  • dPCNA
  • l(2)02448
  • l(2)56F
  • l(2)56Fa
  • l(2)s1534
  • pcna
  • AO090003000846
  • Pcna/cyclin
  • cb16
  • fb36g03
  • wu:fa28e03
  • wu:fb36g03
  • pcna-A
  • Proliferating cell nuclear antigen
  • proliferating cell nuclear antigen
  • DNA polymerase III sliding clamp
  • proliferating cell nuclear antigen L homeolog
  • Proliferating cell nuclear antigen-like
  • proliferating cell nuclear antigen 1
  • PCNA
  • pcna
  • MMP_RS08810
  • ANI_1_512014
  • AOR_1_1500154
  • Pcna
  • pcna.L
  • MGC53867
  • pcna1
RFP tag
Fluorescence Microscopy (FM), Immunofluorescence (IF), Live Cell Imaging (LCI)
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Purpose With the Cell Cycle Chromobody® (CCC) plasmid you can observe cellular processes in response to external stimuli in your cell line of interest.
Brand Chromobody®
Specificity All Chromobody® binding domains are carefully selected to not interfere with endogenous protein function and offer the unique possibility to trace native proteins in living cells.
Characteristics Chromobodies® are a novel species of extremely small intracellular functional antibodies.

Chromobodies® are based on the antigen binding domain (VHH) derived from heavy chain antibodies of camelids genetically fused to a fluorescent protein. As opposed to conventional antibodies, these innovative fluorescent nanoprobes are suitable for real time analyses and can visualize endogenous cellular structures and processes in live cells.
Thus, Chromobodies® are the perfect reagents for cellular research including High-Content Analysis (HCA).

The Chromobody® plasmid allows you to create your own cell lines or transiently express the Chromobody®.
Background Antibodies - extremely powerful tools in biomedical research - are large complex molecules (150 kDa) consisting of two heavy and two light chains. Due to their complex structure the use of antibodies is often limited.

Camelidae (camels, dromedaries, llamas and alpacas) produce functional antibodies devoid of light chains, so called heavy chain antibodies (hcAbs). hcAbs recognize and bind their antigens via a single variable domain (VHH). These VHH domains are the smallest intact antigen binding fragments (molecular weight: 13 kDa), characterized by excellent stability, high specificity and affinities in the low nanomolar range.

PCNA is highly conserved from yeast to man. In eukaryotic cells, PCNA plays a key role in DNA replication, repair, cell cycle regulation, and post-replicative transactions like DNA methylation and chromatin remodelling.or simultaneous read out of Cell size and morphology, Nuclear morphology, Progression of S phase (detailed detection of early-mid-late S phase), Mitosis
Application Notes Chromobodies® are a novel species of intracellular functional antibodies. Chromobodies® are made by genetically fusing the antigen binding domain (VHH) derived from heavy chain antibodies of camelids to fluorescent proteins, e.g. TagGFP2 or TagRFP.

With ChromoTek's unique Chromobody®-Technology the discrepancy and limitations of antibodies and fluorescent protein tagged reporters can now be overcome. You are now able to study the endogenous proteins of your target of interest in living cells - instead of artificial fluorescent fusion proteins which may behave differently from the real protein or may even be toxic to the cells. In combination with High-Throughput Microscopy and automated image analysis Chromobodies® set up a new class of intracellular biomarkers for High-Content Analysis (HCA). Our proof-of-principle studies include:
  • cell cycle monitoring
  • real time apoptosis studies
  • detection of posttranslational modification

These reagents thus significantly expand both the quality and the quantity of information that can be gathered in HCA.

The powerful Cell Cycle Chromobody® enables you to screen for compounds such as cancer drugs for effects on the cell cycle and toxicity in one experiment. You can apply this HCA assay in early drug development and validation as well as in basic research. Cell Cycle Chromobody® visualizes proliferating cell nuclear antigen (PCNA), the homotrimeric ringshaped protein that encircles the DNA and acts as a stationary loading platform for multiple, transiently interacting factors participating in various DNA transactions. As an essential cellular component,

Features of Cell Cycle Chromobody:
Trace dynamic changes during the cell cycle in real time
Monitor the distribution of an endogenous cell cycle marker protein - in comparison to fluorescent fusion proteins no overexpression artifact or cytotoxic effects
Does not affect cellular functions
Also available as stable fluorescent HeLa and U2OS cell lines ready to go for cellular screening.

Protocol New way to trace cell cycle progression in live cells non-invasively!

With the CCC plasmid you obtain the sequence of the Alpaca antibody to PCNA fused to TagRFP or TagGFP. Upon transformation, your cells express the CCC Chromobody® and PCNA is visualized by binding of the fluorescent Chromobody®. The transient binding does not influence PCNA function and thus offers you the unique possibility to observe cell cycle progression in your cells.

Use the CCC to:
Perform dynamic, non-invasive, multi-parameter assays
Multiplex with other procedures, including antibody stainings
Analyze the effect of compounds
Restrictions For Research Use only
Storage -80 °C
Storage Comment Store at -80°C. Short term storage at -20°C.
Product cited in: Burgess, Lorca, Castro: "Quantitative live imaging of endogenous DNA replication in mammalian cells." in: PLoS ONE, Vol. 7, Issue 9, pp. e45726, 2012 (PubMed).