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.
GFP mass spec
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
Best GFP antibody for IP,
Best antibody for immunoprecipitation,
The ChromoTek Histone-Chromobody®* is a novel probe to visualize endogenous histone H2A-H2B heterodimer in live cells:
live cell imaging,
live cell microscopy,
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?
mNeonGreen Western blot,
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.
ChromoTek’s Marketing Manager Dr. Christoph Eckert has been interviewed by Biocompare, the buyers’ guide for life scientists, on labels for super resolution microscopy.
Super resolution microscopy,
Practical considerations and instructions
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:
single domain antibody,
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.
- 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.