ChromoBlog

What is one-step immunostaining?

Posted by Dr. Astrid Sitte on Jul 26, 2019 2:45:28 PM

Simultaneous immunostaining, also called one-step immunostaining vs. sequential immunostaining. Nano-Secondaries stain different primary antibodies equally well in one-step staining and sequential staining.

HeLa cells were immunostained with different primary antibodies and Nano-Secondaries Alexa Fluor® 647 (1:1,000, magenta). Cell nuclei were stained with DAPI (blue). Scale bar, 20 μm.

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Topics: live cell imaging, Secondary antibody, Nano-Secondaries, One-step immunostaining, staining

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

Fluorescent protein tags

Posted by Christian Linke-Winnebeck on May 28, 2019 5:02:54 PM

Introduction

Fluorescent proteins (FPs) have been used as protein tags since the mid-1990s mainly for cell biology and fluorescence microscopy. These tags have not only revolutionized cell biology by enabling the imaging of almost any protein, they are also used in biochemical applications. An important example is the immunoprecipitation and affinity purification of FP-tagged proteins, which was enabled by the development of affinity resins with high yield, purity, and affinity such as ChromoTek’s Nano-Traps (https://www.chromotek.com/products/detail/product-detail/nano-traps/).

In this blog we provide a review of

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Topics: Immunoprecipitation, mCherry, Immunofluorescence, VHH, Nanobody, mNeonGreen, GFP, GFP Nanobody, GFP Antibody, TurboGFP, EGFP, Western blot, SNAP

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

Working with green fluorescent proteins: Tools and properties

Posted by Christoph Eckert on Feb 7, 2019 1:18:14 PM

Jellyfish Green Fluorescent Protein (GFP) and its derivatives are still the most frequently used fluorescent proteins in biomedical research. Recently, additional green fluorescent proteins have been discovered in higher animals such as crustaceans and lancelets. These FPs share a common fold, but diverge widely in their primary sequence. Thus, they require novel, dedicated antibody research tools. Here is an overview about EGFP (the most commonly used GFP derivative), TurboGFP and mNeonGreen.

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Topics: Nanobody, mNeonGreen, mNeonGreen immunoprecipitation, GFP, GFP Immunoprecipitation, TurboGFP, TurboGFP immunoprecipitation, EGFP

ChromoTek’s Nanobody helps LMU scientists in their quest to understand a giant protein complex

Posted by Christian Linke-Winnebeck on Sep 20, 2018 3:42:47 PM

A collaboration between ChromoTek GmbH and Ludwig-Maximilians-University Munich (Germany) has yielded a novel nanobody that sheds new light on an important protein complex called INO80. This work was recently published in the high-impact journal Nature Structural & Molecular Biology (Knoll et al. 2018).

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Topics: Immunoprecipitation, Nanobody, Crystallisation

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

GFP- Nanobodies and RFP- Nanobodies conjugated to quantum dots

Posted by Christoph Eckert on Jul 12, 2018 8:39:33 AM

Chromotek’s GFP- binding protein, an anti- GFP Nanobody, is a very small and effective tool for binding and visualizing GFP-tagged proteins. In their recent publication, Modi et al. successfully functionalized quantum dots with the GFP- Nanobody (QD GFP- Nanobody). Thus, they created a small GFP- specific label with a very strong fluorescent signal.

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Topics: GFP Nanobody, RFP Nanobody, Quantum dots, single particle tracking, Quantum dot Nanobody

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