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

The best anti-GFP antibody for immunoprecipitation: GFP-Trap

Posted by Christoph Eckert on Sep 7, 2017 1:05:34 PM

Life science laboratories apply green fluorescent proteins (GFP) to study protein localization, interaction and dynamics in fluorescence microscopy. Immunoprecipitation (IP), mass spectrometry (MS), co-immunoprecipitation (Co-IP) and/or affinity purification investigate more aspects including posttranslational modifications (PTMs), DNA binding, and protein-protein interaction. Here, we compare two different antibody systems for immunoprecipitation of GFP-fusion proteins: GFP-Trap and anti-GFP IgG antibody

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Topics: GFP, GFP Immunoprecipitation, GFP-Trap, Best GFP antibody for IP, GFP VHH, GFP Nanobody, Best antibody for immunoprecipitation

mNeonGreen: A green fluorescent protein that is not a GFP variant

Posted by Christoph Eckert on Jul 19, 2017 8:20:46 AM

Recently a new bright monomeric yellow-green fluorescent protein has been published, which is called mNeonGreen. This protein has already been frequently used for mainly microscopic applications in both wide-field microscopy and super resolution microscopy. What is mNeonGreen all about?

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Topics: mNeonGreen, mNeonGreen Antibody, mNeonGreen immunofluorescence, mNeonGreen immunoprecipitation, mNeonGreen Western blot, mNeonGreen VHH, GFP, mNeonGreen sequence

New limit in protein complex stability: GFP-binding protein:GFP complex

Posted by Klaus Herick on Feb 3, 2017 3:56:34 PM

Although traditional IgG antibodies are often used for immunoprecipitation and protein interaction analysis, GFP-binding protein (GBP, ChromoTek gt-250) and other ChromoTek VHHs (https://www.chromotek.com/about-us/the-alpaca-antibody-advantage/) easily outcompete those under challenging conditions such as elevated temperatures, varying pH or high denaturant concentrations. In fact, GBP tightly binds GFP-fusion proteins even in 8 M urea at 51 °C, which sets a new limit in protein complex stability, virtually unrivalled by any other capture molecule protein tag pair.

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Topics: GFP-Trap, Nano-Trap, GFP- Binding Protein, GFP

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