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Klaus Herick

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Split fluorescent protein technology

Posted by Klaus Herick on Nov 12, 2019 2:57:15 PM

ChromoTek scientists Michael Metterlein and Christian Linke-Winnebeck have published a whitepaper that provides a comprehensive overview with key developments in the field of split fluorescent protein technology. It also includes a selection of case studies on how ChromoTek’s Nano-Traps have been applied to exploit the full potential of this technology for example in protein-protein interaction studies. Particularly, the ChromoTek GFP-Trap has been successfully applied to multiple assays using different split GFP variants. Assay types include protein self-complementation, bimolecular fluorescence complementation (BiFC), tripartite fluorescence complementation (TriFC), and bimolecular complementation affinity purification (BiCAP).

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Spot Capture and Detection Peptide Tag and Nanobody

Posted by Klaus Herick on Sep 9, 2019 1:37:29 PM

The Spot-Nanobody (green) binds to the Spot-Tag sequence motif PDRVRAVSHWSS. Upon binding, the Spot-Tag peptide is embedded on the surface of the Spot-Nanobody and becomes a β-sheet extension of the Spot-VHH. Defined interactions of the Spot-Nanobody’s side chains to the Spot-peptide determine specificity. In addition, the Spot-peptide is clamped by two amino acid side chains of the Spot-Nanobody. This binding mechanism elucidates why the Spot-Nanobody binds with high affinity to the Spot-Tag.

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Topics: Spot-Tag, Spot Nanobody, Spot-Label

Which are 2018's most used Antibodies?

Posted by Klaus Herick on Aug 26, 2019 4:29:03 PM

 
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Topics: GFP-Trap, GFP Immunoprecipitation, GFP VHH, GFP Nanobody

Important facts to know when using Nano-Secondaries

Posted by Klaus Herick on Jul 23, 2019 1:27:20 PM

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Topics: VHH, Nanobody, Secondary antibody, Nano-Secondaries, Alexa Fluor, One-step immunostaining

GFP-Booster and RFP-Booster conjugated to Alexa Fluor® 647 or conjugated to ATTO647N? That is the question!

Posted by Klaus Herick on Jul 19, 2019 10:12:05 AM

Immunostaining in HeLa cells low expressing Tubulin-GFP.
Left: GFP signal of Tubulin-GFP (green) and DAPI stain (blue);
Right: Tubulin-GFP detection by GFP-Booster coupled to Alexa Fluor 647

We currently offer our GFP- and RFP-Booster conjugated to two different far-red dyes:

Alexa Fluor® 647 and  ATTO647N.

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Topics: GFP-Booster, RFP-Booster, Nanobody, Alexa Fluor

Why are recombinant Nanobodies/ VHHs beneficial?

Posted by Klaus Herick on Jun 13, 2019 4:00:00 PM

Nanobodies are the binding domains of heavy chain only antibodies from Camelids. Nanobodies can be recombinantly produced in bacterial and other animal-free expression systems depending on the actual Nanobody construct. In contrast, classical IgG antibodies are composed of two heavy chains and two light chains and are traditionally produced using hybridoma technlogies or are isolated from a host’s blood.

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Topics: Nanobody, Secondary antibody, Validation, recombinat expression

Advantages of recombinant Nano-Secondaries

Posted by Klaus Herick on May 23, 2019 4:10:12 PM

Nano-Secondaries are monoclonal Nanobodies that bind to primary antibodies in a species and subtype specific manner, i.e. Nano-Secondaries are very precise secondary antibodies. They are recombinantly produced in bacterial or other animal-free expression systems; in contrast, classical mono- or polyclonal secondary antibodies are traditionally produced using hybridoma technologies or are isolated from a host’s blood.

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Topics: Secondary antibody, Validation, recombinat expression, Nano-Secondaries

22 References about Myc-Trap

Posted by Klaus Herick on Sep 6, 2018 4:29:17 PM

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.

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Topics: Nano-Traps, Myc-tag IP, Myc Nanobody, Myc-Trap

First Peptide-tag specific Nanobody applied in super resolution microscopy (SRM)

Posted by Klaus Herick on Mar 2, 2018 11:02:00 AM

You may have noticed the recent publication “A peptide tag-specific nanobody enables high-quality labeling for STORM imaging” of Virant et al (2018) in Nature Communications doi: 10.1038/s41467-018-03191-2, where for the first time a peptide-tag specific Nanobody was applied in dSTORM imaging: The authors have described and discussed the performance of a BC2 peptide-tag specific Nanobody to image BC2 peptide tag fusion proteins in super resolution microscopy, i.e. dSTORM. This Blog features that BC2 peptide-tag specific Nanobody, which ChromoTek markets under the names Spot-Label® and Spot-Trap®. In addition, this Blog figures the significantly improved version of the BC2 peptide -tag, which is called Spot-Tag®.

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Topics: Nanobody, Spot-Tag, Capture/detection System, Spot Nanobody, Spot-Trap, Spot-Label, Spot super resolution microscopy, Spot STED, Spot immunofluorescence, BC2tag

Affinity purification, depletion, and immunoprecipitation of GST-tagged proteins

Posted by Klaus Herick on Jul 6, 2017 1:30:48 PM

Multiple literature references for GST-Trap


This article provides an overview of scientific publications that reference the ChromoTek GST-Trap superior binding performance for various applications. These include immunoprecipitation (IP), Co-IP of Glutathione S-Transferase (GST)-fusion proteins, and Luminex bead assays of GST-tagged and GFP-tagged proteins. Besides, usages for depletion of GST (glutathione sepharose)  after cleavage from recombinant fusion proteins and affinity purification of GST-tagged proteins are possible.

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Topics: Immunoprecipitation, GST, affinity resin, GST immunoprecipitation, references, affinity purification, GST pulldown

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