ChromoTek Cell Cycle Chromobody® plasmid (TagRFP)

DNA plasmid encoding for anti-PCNA VHH (anti-PCNA Nanobody) fused to fluorescent protein TagRFP. PCNA is a cell cycle marker.

Specificity

PCNA

Applications

IF, Live cell imaging

Vector type

Mammalian expression vector

Reporter

TagRFP

Publications

16

Cat no : ccr

Synonyms

Chromobody, PCNA Chromobody

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Validation Data Gallery

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  Live cell imaging of HeLa cells: Live cell imaging of HeLa cells stable expressing Cell Cycle-Chromobody (TagRFP). Images from a time series of 22 hours are shown

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  Cell Cycle-Chromobody signal during the cell cycle: In G1, the Chromobody signal is homogeneously distributed throughout the nucleus and cytoplasm. During S phase it accumulates in the nucleus and visualizes the formation of replication foci. In G2, the foci disappear and the cell divides (mitosis).

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  Time lapse video: Live cell imaging of the stable cell line HeLa-Cell Cycle Chromobody TagRFP expressing Cell Cycle-Chromobody.

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Product Information

DNA plasmid encoding for anti-PCNA VHH (anti-PCNA Nanobody) fused to fluorescent protein TagRFP. PCNA is a cell cycle marker.

• Description: The Cell Cycle-Chromobody allows visualization of the endogenous cell cycle marker protein PCNA:

  • Trace dynamic changes during the cell cycle in real-time

  • Monitor the distribution of an endogenous cell cycle marker protein without artifacts or cytotoxic effects when overexpressing fluorescent fusion proteins

  • Does not affect cellular functions

• Applications: Live cell imaging; Visualizing endogenous cell cycle marker protein PCNA; Actin as biomarker or control
• Specificity: PCNA (proliferating cell nuclear antigen), tested in human cells and zebrafish
• Reporter: TagRFP
• Vector type: Mammalian expression vector
• Promoter: Constitutive CMV IE
• Codon usage: Mammalian
• Selection: Kan/Neo
• Sequence: With the Cell Cycle-Chromobody plasmid you receive the sequence information of the Alpaca antibody to PCNA fused to TagRFP, as well as the full vector sequence.
• Transfection: Transfection of Chromobody plasmids into mammalian cells can be done with standard DNA-transfection methods, e.g. lipofection (Lipofectamine 2000® from Thermo Scientific), according to the manufacturer’s protocol for the transfection reagent. Please choose the transfection method that works the best for your cell type.
• Microscopy techniques: Wide-field epifluorescence microscopy; confocal microscopy; super-resolution microscopy e.g. STED and 3D-SIM; high-content microscopy and analysis
• Storage Condition: Shipped at ambient temperature. Store at -20 °C

Publications

Application	Title
	Nature; Equilibrium between nascent and parental MCM proteins protects replicating genomes.; Authors - Hana Sedlackova
	Nature; CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition.; Authors - David Gallo
	Nat Cell Biol; Nuclear F-actin counteracts nuclear deformation and promotes fork repair during replication stress.; Authors - Noa Lamm
	Nat Cell Biol; 53BP1 nuclear bodies enforce replication timing at under-replicated DNA to limit heritable DNA damage.; Authors - Julian Spies
IHC	Mol Cell; DNA Replication Determines Timing of Mitosis by Restricting CDK1 and PLK1 Activation.; Authors - Bennie Lemmens
	EMBO J; Evidence for the nuclear import of histones H3.1 and H4 as monomers.; Authors - Michael James Apta-Smith