ChromoBlog

Kourosh Zolghadr

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Chromobody® plasmids: Good things come to those who wait!

Posted by Kourosh Zolghadr on Jun 5, 2014 4:50:00 PM

This time last spring we started marketing our Chromobody® plasmids. Our customers realized the impact of the technology and were immediately beginning to work with the Chromobodies®. Now, one year later the first papers are being published demonstrating the broad applicability of this intracellular fluorescent antibody technology: 

The first is a paper in Molecular Cell by the Lindqvist Lab at the Karolinska Institutet in Stockholm, Sweden. They developed a FRET-based system for accurate quantification of fluorescence from single cells. By visualizing endogenous PCNA at replication foci with the Cell Cycle Chromobody® they nicely show that the mitotic entry network is linked to the completion of S phase. It does not depend on protein accumulation through G2 but is activated by mitotic phosphorylations at the end of S phase. The method they present allows analyzing live-cell as well as extracting temporal information from fixed cells based on endogenous marker proteins. (Akopyan, K.; Silva Cascales, H.; Hukasova, E., et al. Assessing kinetics from fixed cells reveals activation of the mitotic entry network at the S/G2 transition. Molecular cell. 2014, 53, 843-853. http://dx.doi.org/10.1016/j.molcel.2014.01.031

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Topics: Chromobodies, Cell Cycle Chromobody, Chromobody, Chromobody plasmid, Actin Chromobody, Lamin Chromobody, Dnmt1 Chromobody, custom Chromobody, intracellular antibody, PCNA

New Nano-Trap: Focus on MK2!

Posted by Kourosh Zolghadr on May 13, 2014 4:09:00 PM

MAP kinase-activated protein kinase 2 (MAPKAP-K2, MK2; Gene ID: 9261) is a 400 AA (46kDa) large enzyme that plays a central role mainly in the inflammatory response and cytokines production. It belongs to the serine/threonine-protein kinase family and is also involved in endocytosis, reorganization of the cytoskeleton, cell migration, cell cycle control, chromatin remodeling, DNA damage response and transcriptional regulation.1-4

Following stress, it is phosphorylated (at Thr-222, Ser-272 and Thr-334) and activated by MAP kinase p38-alpha/MAPK14, leading to phosphorylation of substrates.5 Phosphorylation of Thr-334 (located between the kinase domain and the C-terminal regulatory domain) may serve as a switch for MK2 nuclear import and export. Phosphorylated MK2 masks the nuclear localization signal at its C-terminus by binding to p38. It unmasks the nuclear export signal, which is part of the second C-terminal helix packed along the surface of kinase domain C-lobe, and thereby carries p38 to the cytoplasm.6, 7

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Topics: Nano-Trap, MK2-Trap, Immunoprecipitation, Co-IP

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