ChromoBlog

1. What is GFP?

Nanobody-Fc fusion - An alternative Nanobody format that combines its special epitope binding properties with the detection options for Fc-domains

What are heavy chain antibodies?

When time in the lab is limited, learn how to save precious hours on your immunoprecipitation, immunofluorescence and western blotting experiments.

Due to the current situation with the COVID-19 pandemic, many researchers around the world are faced with restrictions on time and resources in the lab. As we adapt to these changes, the pressure is on to find ways to save time without compromising on results. Proteintech and ChromoTek offer solutions to save you hours on your current immunoassay protocols, helping you achieve publication worthy results in a fast and reliable manner.

An extract of the published literature since 2016

This blog provides references from the last 4 years in virus research using GFP. Most publications report how immunoprecipitation (IP)/Co-IP of GFP-fusions was conducted to identify host cell binding partners of virus proteins. In addition, mass spectrometry analysis and functional assays have been performed.

New iST Nano-Trap kits for immunoprecipitation (IP) and sample preparation for mass spectrometry (MS) in just 4 easy steps:

Crystal structure of the anti-GFP VHH-Green Fluorescent Protein complex.
The GFP Nanobody is displayed blue and the GFP in green color.

UV crosslinking techniques are the method of choice for a comprehensive analysis of in-vivo-mRNA targets of an RNA-binding protein (RBP). In the recent publication of Olgeiser et al. (2019), the authors applied individual-nucleotide resolution UV crosslinking and immunoprecipitation (iCLIP) to study fungal mRNA transport. For this approach, they have used strains expressing GFP-tagged versions of the two RBPs Grp1 and Rrm4; an optimized protocol was developed to uncover that Grp1 and Rrm4 conjointly bind thousands of shared target messenger ribonucleoproteins (mRNPs) in the fungus U. maydis. The protein:RNA complexes were immunoprecipitated in a multiple detergent containing buffer using ChromoTek’s GFP-Trap Magnetic Agarose. This is a transcriptome‐wide view to an endosomal mRNA transport machinery.

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

• green fluorescent proteins
• red fluorescent proteins
• self-labelling proteins, that require the covalent coupling of a fluorescent molecule

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.

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