ChromoBlog

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

Visualize Histones in live cells: Histone-Chromobody

Posted by Christoph Eckert on Aug 3, 2017 1:22:32 PM

The ChromoTek Histone-Chromobody®* is a novel probe to visualize endogenous histone H2A-H2B heterodimer in live cells:

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Topics: live cell imaging, Histone Chromobody, live cell microscopy, Nucleosome, Chromatin Marker, Histone

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 Western blot, mNeonGreen VHH, mNeonGreen immunoprecipitation, mNeonGreen immunofluorescence, GFP, mNeonGreen sequence

New anti-mNeonGreen antibody & Nano-Trap for Immunofluorescence, Western Blot & Immunoprecipitation

Posted by Christoph Eckert on Jul 13, 2017 11:29:11 AM

ChromoTek is proud to introduce new tools for mNeonGreen fusion proteins: 

Click here and request a free sample:

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Topics: mNeonGreen, mNeonGreen Antibody, mNeonGreen Western blot, mNeonGreen VHH, mNeonGreen-Trap, mNeonGreen immunoprecipitation, mNeonGreen immunofluorescence

Affinity purification, depletion, and immunoprecipitation of GST-tagged proteins

Posted by Klaus Herick on Jul 6, 2017 1:30:48 PM

Multiple literature references for GST-Trap


This article provides an overview of scientific publications that reference the ChromoTek GST-Trap superior binding performance for various applications. These include immunoprecipitation (IP), Co-IP of Glutathione S-Transferase (GST)-fusion proteins, and Luminex bead assays of GST-tagged and GFP-tagged proteins. Besides, usages for depletion of GST (glutathione sepharose)  after cleavage from recombinant fusion proteins and affinity purification of GST-tagged proteins are possible.

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Topics: Immunoprecipitation, GST, affinity resin, GST immunoprecipitation, references, affinity purification, GST pulldown

ChromoTek Nano-Booster for super resolution microscopy is featured at Biocompare

Posted by Klaus Herick on Jun 2, 2017 4:21:49 PM

ChromoTek’s Marketing Manager Dr. Christoph Eckert has been interviewed by Biocompare, the buyers’ guide for life scientists, on labels for super resolution microscopy.

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Topics: GFP-Booster, RFP-Booster, Super resolution microscopy, VHH, Nanobody

Myc-Trap for both immunoprecipitation and affinity purification of Myc-tagged proteins

Posted by Klaus Herick on Jun 2, 2017 11:41:33 AM

Practical considerations and instructions

Introduction

The ChromoTek Myc-Trap is an affinity resin that consists of Myc-binding protein derived from an Alpaca single domain antibody, also called anti-Myc VHH or “anti-Myc nanobody”, which is coupled to agarose and magnetic agarose beads. The Myc-Trap provides advantages over conventional IgG antibodies when applied in immunoprecipitation (IP) and affinity purification experiments:

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Topics: Immunoprecipitation, Myc-tag, Myc-tag IP, single domain antibody, Myc Nanobody

Immunoprecipitation of Myc-tagged proteins – How it works

Posted by Christoph Eckert on May 26, 2017 1:57:07 PM

ChromoTek Myc-Trap® is an effectiveness optimized tool for immunoprecipitation (IP/Co-IP) and affinity purification of Myc-tagged proteins. Depending on whether you use a single Myc-tag, i.e. 1xMyc (EQKLISEEDL) or a double Myc-tag fused to the protein of interest, i.e. 2xMyc (EQKLISEEDLEQKLISEEDL) you can adjust your experiment. Here, we introduce the epitope of Myc-Trap to discuss its performance for afore mentioned applications. 

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Topics: Immunoprecipitation, Myc-tag, Myc-tag IP

Analyze protein interactions live in mammalian cells

Posted by Christoph Eckert on May 16, 2017 4:18:31 PM
  • Do you have GFP or RFP protein constructs and/or libraries?
  • Do you want to screen agonists or antagonists?
  • Do you want to save time and start your experiment without method set-up?

Just do it. All you need is the ChromoTek F2H Kit Basic: Transfect the bait and the prey plasmids with the Platform Reagent into the F2H cells. Image and quantify your results already at the next day.

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Topics: F2H assay, PPI

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