ChromoBlog

GFP and RFP-Booster for better immunofluorescence imaging

Posted by Christoph Eckert on Jan 29, 2020 5:37:07 PM

GFP-Booster and RFP-Booster conjugated to Alexa Fluor® 488, 568 and 647:

  • Stronger signal in confocal and standard fluorescence microscopy
  • More options in super-resolution microscopy including STORM and MINFLUX
  • Constant degree of labeling (DOL = 2 dyes per Booster) for higher resolution
  • More than 95% labeling efficiency
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Topics: GFP-Booster, RFP-Booster, GFP Nanobody, RFP Nanobody, EGFP

What GFP-Trap should I use for my immunoprecipitation?

Posted by Dr. Astrid Sitte on Jan 20, 2020 4:42:38 PM

ChromoTek GFP-Trap® is optimized for the immunoprecipitation of GFP-tagged proteins and their interacting factors.

The GFP-Trap consists of the ChromoTek anti-GFP Nanobody/ VHH that is coupled to 3 different types of beads or is immobilized in a 96 multiwell plate:

  • Agarose beads
  • Magnetic Agarose beads
  • Dynabeads™
  • 96 Multiwell Plate

Immunoprecipitation with GFP-Trap.
I: Input, FT: Flow-Through, B: Bound

Based on the properties of the different matrices (see table), we recommend:

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Topics: GFP-Trap, GFP Immunoprecipitation, GFP Nanobody, Dynabeads

GFP-Trap Dynabeads: Most cited GFP-Nanobody conjugated to Dynabeads

Posted by Christoph Eckert on Jan 17, 2020 11:27:30 AM

GFP-Trap® Dynabeads is optimized for the immunoprecipitation (IP) of large GFP-tagged proteins and for Co-IP of protein complexes. It consists of ChromoTek’s established anti-GFP Nanobody conjugated to Dynabeads™. Hence, also GFP-Trap Dynabeads has the very high affinity of 1 pM like GFP-Trap Agarose and Magnetic Agarose.

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Topics: GFP Immunoprecipitation, GFP Nanobody, Dynabeads

Which are 2018's most used Antibodies?

Posted by Klaus Herick on Aug 26, 2019 4:29:03 PM

 
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Topics: GFP-Trap, GFP Immunoprecipitation, GFP VHH, GFP Nanobody

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/).

In this blog we provide a review of

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

GFP- Nanobodies and RFP- Nanobodies conjugated to quantum dots

Posted by Christoph Eckert on Jul 12, 2018 8:39:33 AM

Chromotek’s GFP- binding protein, an anti- GFP Nanobody, is a very small and effective tool for binding and visualizing GFP-tagged proteins. In their recent publication, Modi et al. successfully functionalized quantum dots with the GFP- Nanobody (QD GFP- Nanobody). Thus, they created a small GFP- specific label with a very strong fluorescent signal.

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Topics: GFP Nanobody, RFP Nanobody, Quantum dots, single particle tracking, Quantum dot Nanobody

On-bead digestion protocol: From immunoprecipitation with nanobodies to MS analysis

Posted by Christoph Eckert on Sep 15, 2017 3:52:48 PM

What makes on-bead digestion favorable?

Just pull down your protein of interest with immobilized nanobodies, also termed VHHs or single domain antibodies. Then follow the on-bead digestion protocol (see below) and submit the digest to your core facility for effective mass spectrometer analysis of (co-) precipitated proteins.

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Topics: GFP-Trap, GFP Immunoprecipitation, GFP VHH, GFP Nanobody, on-bead digestion, GFP mass spec

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

Nano-Booster FAQs

Posted by Larisa Yurlova on Feb 19, 2016 2:48:05 PM

ChromoTek Nano-Boosters are ideal for Super-Resolution and “traditional” fluorescence microscopy because of their high affinity and extremely small size of just 2 to 3 nm. Technically speaking, the GFP-and RFP-Boosters are composed of the highly specific GFP- or RFP-binding domains of alpaca antibodies (also called “nanobodies”), covalently coupled to a selection of fluorescent dyes.

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Topics: Nano-Booster, GFP-Booster, RFP-Booster, GFP Nanobody, RFP Nanobody, mCherry Nanobody

ChromoTek's Nanobodies are versatile and proven affinity tools for multiplexed analysis of protein-protein interactions using Luminex® or Biacore™ assay technologies

Posted by Laurentia Tanase on Aug 14, 2015 11:56:12 AM

Benefit from high binding affinities and extraordinary functional stabilities of Nanobodies

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Topics: GFP-Trap, GFP Nanobody, GST Nanobody

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