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Christian Linke-Winnebeck

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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/).

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

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

5 Tips for better immunoprecipitation (IP)

Posted by Christian Linke-Winnebeck on Jul 21, 2016 1:10:15 PM

Pull-down of proteins can be difficult, particularly when they are expressed at low levels. Here we present 5 tips, which will considerably improve your IP results.

 

  1. Protein concentration matters

The higher the protein concentration, the higher the IP yield. Try to use a concentration as high as possible.It makes a significant difference if the concentration of your protein during IP is 1 nM (low protein expression level), about 50 nM (intracellular endogenous protein expression level, will be diluted by buffer for IP) or 1000 nM (high protein concentration).

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Topics: Nano-Trap, Immunoprecipitation, Co-IP, GST pulldown, MBP pulldown

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