ChromoBlog

Since the discovery of the green fluorescent protein (GFP) it has been widely applied in fluorescence microscopy as a tool to study proteins in their native cellular environment. However, GFP has several intrinsic limitations, such as low signal intensity, fast photo bleaching and signal loss after chemical treatment. The GFP-Booster, which is composed of an anti-GFP Nanobody conjugated to a fluorescent dye, enhances, stabilizes, and re-establishes the signal of GFP fusion proteins in immunofluorescence. In this blog, we answer why the small size of the GFP-Booster is of advantage compared to regular antibodies in immunofluorescence and provide an overview of the GFP variants bound by the GFP-Booster as well as the available dyes.

The high stability of the GFP-Trap^® enables its use in virtually any lysis and wash buffer. Therefore, it is possible to remove unwanted proteins, reduce background in your IP, or apply the GFP-Trap in applications requiring harsh buffer conditions. The GFP-Trap bound to the GFP-fusion protein can, for example, be used in ubiquitination assays or in the presence of Urea, which is used for the total inactivation of any phosphatase activity in Co-IP/MS for phosphorylation studies.

Flag®-tag (or DYKDDDDK-tag) is a commonly used short peptide tag for multiple applications such as immunoprecipitation (IP), protein purification, immunofluorescence, and Western blotting (WB). In this blog, we provide an introduction to the IP of Flag®-tagged proteins from cellular extracts. Using our DYKDDDDK Fab-Trap™ as an example, we elaborate on the different steps of IP and highlight the controls that help you achieve the best result in your experiment.

1. What is Flag-Tag?

1. What is GFP?

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Myc-tag is a peptide tag derived from the c-Myc protein. The Myc-tag can be used for many capture and detection applications such as immunoprecipitation, immunofluorescence and protein purification.

V5 tag is a short peptide tag for detection and purification of proteins. The V5 tag can be fused/cloned to a recombinant protein and detected in ELISA, flow cytometry, immunoprecipitation, immunofluorescence, and Western blotting with antibodies and Nanobodies.

SNAP-tag and CLIP-tag are self-labeling protein tags. The SNAP-tag protein and the CLIP-tag protein can be fused to a protein of interest (POI) and used for cellular and biochemical analysis.

HaloTag is a protein tag. The HaloTag protein can be fused to your protein of interest (POI) which enables it for cellular and biochemical analysis. It can also be used for live cell imaging.