Different antibody formats can be used in IP applications, including full-length antibodies, Fab-fragments, and Nanobodies. ChromoTek’s Fab-Trap™ is an IP reagent based on an antibody Fab-fragment. In this blog, we elaborate on the basic principle and advantages of Fab-fragment based reagents for IP. More simply, we ask “Why is the Fab-Trap™ Fab-ulous?”
Immunoprecipitation (IP) is a technique used to isolate a protein from a cellular extract. The protein of interest (POI) is recognized by a specific antibody (Ab) conjugated to beads. Different antibody formats can be used in IP, e.g., a full-length Ab, a Fab-fragment, or a Nanobody. Below, we discuss the advantages and limitations of the different Ab formats.
We are happy to present an updated list of scientific publications that have been published based on data generated by Myc-Trap. See detailed list below.
See the advantages of ChromoTek’s RFP-Trap in ChIP applications as demonstrated by Dr. Caroline Rivers, Laboratory for Integrative Neuroscience & Endocrinology, University of Bristol. In absence of suitable antibodies for an inducible hormone receptor, mCherry has been fused to the N-terminus of the receptor. In order to effectively pull down the mCherry fusion protein DNA complex, ChromoTek's RFP-Trap was used.
It was discovered 35 years ago, was awarded the title „molecule of the year“ in 1993 by the journal Science, and has amassed a whopping 72,000 research papers to date. Hardly any other protein has stirred up so much scientific interest as the transcription factor with the rather unspectacular name p53, which became famous as a tumor suppressor (1). It patrols the nucleus for damaged DNA and prevents cells from dividing or, if the damage is too large, induces apoptosis. Thus, p53 can be regarded as a kind of emergency brake that is pulled when something goes utterly wrong. Somewhat more poetically, Sir David P. Lane, probably the best known researcher in this field and the second most cited UK-based scientist in the 1990’s, called it the „guardian of the genome“ (2).
Chromobodies® are new intracellular functional antibodies visualising dynamic changes of target proteins in live cells. With the help of NMI`s outstanding lentiviral transduction technology, Chromobodies® will be introduced into difficult-to-transfect cell lines such as primary cells. The cooperation will establish innovative cell lines available for phenotypic screening, target validation and high content analysis (HCA). The unique combination of Chromobody® nanoprobes and a comprehensive collection of disease-relevant cellular test systems will lift preclinical compound screening and pathway analysis in the early drug discovery process to a new level of performance.
Benefit from great introductory pricing of our new partner, Bulldog Bio!
ChromoTek GmbH (Munich, Germany) has announced today the signing of a license agreement granting Bayer Pharma AG (Bayer) the right to use ChromoTek's proprietary Cell Cycle Chromobody® Technology for drug profiling by High Content Analysis. Under the terms of the agreement, Bayer will apply Chromobody® technology for real time monitoring of cell cycle progression in drug discovery. Financial terms of the agreement were not disclosed. This agreement follows a collaboration in which ChromoTek provided custom cell line development.
ChromoTek's Chromobody® Technology is a patented, universal live cell imaging technology for high content analysis. Chromobodies are single domain antibodies that have been genetically fused to fluorescent proteins to serve as functional nanoprobes in living cells. The Chromobody® technology allows for the first time to trace endogenous intracellular antigens and to visualize dynamic changes of these targets within living cells. Currently available Chromobodies trace important cellular processes like cell cycle, apoptosis, DNA damage and repair or allow cytoskeletal analysis. The Cell Cycle Chromobody® cell line licensed by Bayer traces the complete cell cycle in real time in live cells in a non-invasive manner while providing full flexibility to multiplex with other procedures.
Dr. Kourosh Zolghadr, Head of R&D at ChromoTek comments: "We are pleased to offer our customers efficient tools for achieving more detailed and physiologically relevant information on the effects of drug candidates on cell cycle progression."
ChromoTek products set new benchmarks for cellular research. Established in 2008 as a spin-off from Munich’s Ludwig Maximilian University and located in Martinsried, Germany’s leading biotech cluster, ChromoTek develops and markets Chromobody® based live cell assays for drug screening and target validation. Other products include immunological and bioimaging reagents such as the GFP- Trap® for the rapid pull-down of GFP fusion proteins, GFP and RFP-Booster to amplify fluorescence signals of GFP or RFP fusion proteins. ChromoTek’s newly developed fluorescence based protein protein interaction assay called Fluorescent-2-Hybrid (F2H® assay) is available as a development service. More than 2.800 customers from all over the world trust in ChromoTek products.
Posted by Katrin on Mar 18, 2013
ChromoTek GmbH (Munich, Germany) offers nanobody based real time assays to visualize and measure the action of candidate drugs or other substances of interest on important regulatory processes in living cells. Presently, four Chromobody® plasmids are available for analyzing the cell cycle, apoptosis, DNA methylation and cytoskeleton dynamics.