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

The importance of blocking when using Nano-Secondaries for IF

Posted by Dr. Astrid Sitte on Jan 28, 2020 5:04:30 PM

Introduction

Blocking is an essential step during the preparation of a sample for immunofluorescence detection. Blocking improves the sensitivity by reducing nonspecific background and therefore increases image quality. Insufficient blocking results in higher background noise and over-blocking can even mask the specific signal.

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Topics: Nano-Secondaries, staining

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:

  • Agarose beads
  • Magnetic Agarose beads
  • Dynabeads™

Based on the properties of the different beads (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

How to overcome the limitations of antibody-based affinity resins for protein purification

Posted by Dr. Astrid Sitte on Dec 18, 2019 2:19:42 PM

Introduction

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Topics: affinity resin, affinity purification, Spot-Tag, Spot Nanobody, protein purification, epitope tag

Tags for affinity purification

Posted by Christoph Eckert on Nov 18, 2019 2:56:52 PM

Affinity tags are very useful tools for protein purification. Fused to the protein of interest, they streamline the purification process by binding to a tag-specific resin. Obviously, tag selection is an important step as the purification tag can affect expression level, solubility, facilitate correct folding, protect from proteolysis, and re-direct proteins to a cellular compartment. In addition, the purification tag determines the affinity resin used.

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Split fluorescent protein technology

Posted by Klaus Herick on Nov 12, 2019 2:57:15 PM

ChromoTek scientists Michael Metterlein and Christian Linke-Winnebeck have published a whitepaper that provides a comprehensive overview with key developments in the field of split fluorescent protein technology. It also includes a selection of case studies on how ChromoTek’s Nano-Traps have been applied to exploit the full potential of this technology for example in protein-protein interaction studies. Particularly, the ChromoTek GFP-Trap has been successfully applied to multiple assays using different split GFP variants. Assay types include protein self-complementation, bimolecular fluorescence complementation (BiFC), tripartite fluorescence complementation (TriFC), and bimolecular complementation affinity purification (BiCAP).

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Bispecific T cell engagers

Posted by Christoph Eckert on Sep 13, 2019 1:04:52 PM
Bispecific T cell engagers are bispecific antibodies designed to link T cells with cancer cells. The simultaneous binding of the bispecific antibody to T cell and tumor cell initiates the activation of the T cell. For this purpose, the bispecific antibody carries two antigen binding domains against the T cell (often CD3 T cell receptor) and against a cell surface protein of the tumor cell.

ChromoTek offers two bispecific T cell engagers to beta-testers:

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Topics: VHH, Nanobody, Bispecific, T cell

Spot Capture and Detection Peptide Tag and Nanobody

Posted by Klaus Herick on Sep 9, 2019 1:37:29 PM

The Spot-Nanobody (green) binds to the Spot-Tag sequence motif PDRVRAVSHWSS. Upon binding, the Spot-Tag peptide is embedded on the surface of the Spot-Nanobody and becomes a β-sheet extension of the Spot-VHH. Defined interactions of the Spot-Nanobody’s side chains to the Spot-peptide determine specificity. In addition, the Spot-peptide is clamped by two amino acid side chains of the Spot-Nanobody. This binding mechanism elucidates why the Spot-Nanobody binds with high affinity to the Spot-Tag.

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Topics: Spot-Tag, Spot Nanobody, Spot-Label

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

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