GFP-Booster: GFP Nanobody for better images in immunofluorescence

Posted by Christoph Eckert on Jan 31, 2022 1:06:06 PM

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.

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Topics: GFP-Booster

GFP-Trap for immunoprecipitation (IP) – stringent washing

Posted by Christoph Eckert on Jan 27, 2022 12:07:53 PM

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.

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How to immunoprecipitate Flag®-tagged proteins

Posted by Christoph Eckert on Dec 16, 2021 10:33:31 AM

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.

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Topics: Immunoprecipitation

Why is the Fab-Trap™ so Fab-ulous?

Posted by Dr. Bennet Reiter on Nov 25, 2021 9:11:20 AM

Different antibody formats can be used in IP applications, including full-length antibodies, Fab-fragments, and Nanobodies. ChromoTek’s Fab-Trap™ is an IP reagent based on an antibody Fab-fragment. In this blog, we elaborate on the basic principle and advantages of Fab-fragment based reagents for IP. More simply, we ask “Why is the Fab-Trap™ Fab-ulous?”

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Topics: Nano-Traps, Immunoprecipitation, Fab-Trap

Advantages and limitations of different antibody formats in immunoprecipitation

Posted by Dr. Bennet Reiter on Nov 25, 2021 8:52:38 AM

Immunoprecipitation (IP) is a technique used to isolate a protein from a cellular extract. The protein of interest (POI) is recognized by a specific antibody (Ab) conjugated to beads. Different antibody formats can be used in IP, e.g., a full-length Ab, a Fab-fragment, or a Nanobody. Below, we discuss the advantages and limitations of the different Ab formats.

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Topics: Nano-Traps, Immunoprecipitation, Nanobody, Fab-Trap

How Nanobodies could advance Sars-CoV diagnostics

Posted by guest authors Teresa Wagner and Ulrich Rothbauer on Oct 15, 2021 11:30:47 AM

ChromoTek and Proteintech recently launched Nanobodies against the SARS-CoV-2 virus. These Nanobody clones were originally developed by the group of Ulrich Rothbauer at the Natural and Medical Sciences Institute, Reutlingen, Germany. In our guest blog, Teresa Wagner and Ulrich Rothbauer discuss the development of these Nanobodies and why Nanobodies are versatile alternatives to classical antibodies in SARS-CoV-2 research.

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Topics: Nanobody

Co-immunoprecipitation troubleshooting

Posted by Dr. Astrid Sitte on Aug 27, 2021 5:21:16 PM

Co-immunoprecipitation (Co-IP) describes the isolation of a protein and its binding partners from a cell extract using a Nanobody or antibody that is bound to beads. The protein, that directly interacts with the Nanobody, or antibody beads is called “bait”. The binding partner that is indirectly precipitated is called “prey”.

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Topics: GFP-Trap, Immunoprecipitation, Co-IP

How to conduct a Co-immunoprecipitation (Co-IP)?

Posted by Dr. Astrid Sitte on Jul 30, 2021 2:39:52 PM

Co-IP describes the immunoprecipitation of a protein of interest and its interacting partners.

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Topics: GFP-Trap, Immunoprecipitation, Co-IP

Flag-tag and 3x Flag-tag: An epitope tag for capture and detection experiments

Posted by Christoph Eckert on Jun 25, 2021 10:35:07 AM

1. What is Flag-Tag?

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Topics: Myc-tag, Spot-Tag, HA-Tag, Flag-Tag

GFP (green fluorescent protein): Properties, origin, specifications, tips

Posted by Christoph Eckert on May 25, 2021 5:13:30 PM

1. What is GFP?

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Topics: GFP, GFP VHH, GFP Nanobody, EGFP

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