Application notes

Chromatin immunoprecipitation (ChIP)

ChIP protocol for A. thaliana using GFP-Trap

The protocol can be applied for all kinds of A. thaliana starting material such as seedlings, leaves and inflorescences.

ChIP protocol for A. thaliana using GFP-Trap

ChIP with RFP-tagged proteins

RFP-Trap for ChIP analysis in transiently expressing a mCherry fusion in mammalian cells.

ChIP with RFP-tagged proteins

CRISPR

Spot-System for CRISPR’ed Spot-Tag

Spot-Trap and GFP-Trap for capture and Spot-Label for detection of CRISPR’ed Spot-Tag and EGFP.

Spot-System for CRISPR’ed Spot-Tag

Elution of GFP

Elution of GFP-fusion protein from the GFP-Trap

The GFP-Trap has a very high affinity and specificity. The GFP-Trap enables very effective pulldowns, but requires special protocols for elution of bound GFP-fusion proteins if they are not analyzed on-bead.

Elution of GFP-fusion protein from the GFP-Trap

Enzyme activity assay

One step purification of GFP-tagged enzymes for enzymatic activity tests

Using the GFP-Trap, GFP-fusion proteins of interest can be efficiently immunoprecipitated from cellular extracts for subsequent assays.

One step purification of GFP-tagged enzymes for enzymatic activity tests

Immunoprecipitation

GFP immunoprecipitation of Arabidopsis thaliana plant samples

This Application Note provides a short protocol for the use of GFP-Trap® Agarose for IPs from Arabidopsis thaliana samples. It is based on more than 70 peer reviewed publications. Please note that the protocol may also serve as inspiration when working with other plants e.g. Nicotiana .

How to do GFP immunoprecipitation of Arabidopsis thaliana plant samples with GFP-Trap

 

Sample preparation of immunoprecipitated EGFP-tagged PARP1 to identify protein-protein interactions

We combined the GFP-Trap® with the fast, sensitive and robust iST sample preparation kit (PreOmics GmbH). The combination of both workflows enables a streamlined sample preparation of immunoprecipitated GFP-fusion proteins for subsequent analysis by mass spectrometry.

Application note

Immunostaining

One-step staining with Nano-Secondaries

Nano-Secondaries can be simultaneously incubated with the primary antibody. One-step immunostaining saves incubation time and reduces washing steps, and hands-on time.

One-step Immunostaining with Nano-Secondaries

 

Blocking in immunofluorescence

What blocking solution should I use for Nano-Secondaries? Blocking increases image quality and therefore, it is an essential step during the preparation of a sample for immunofluorescence detection.

Blocking recommendation for immunostaining with Nano-Secondaries

Mass spectrometry

On-bead digest protocol for mass spectrometry

On-bead digest protocol for mass spectrometry following immunoprecipitation with GFP-Trap, RFP-Trap, Myc-Trap, Spot-Trap, etc..

On-bead digest protocol for mass spectrometry

iST GFP-Trap Kit for mass spectrometry (application note)

GFP-Trap for Co-IP of GFP-fusion proteins plus their interaction partners and iST technology for sample preparation for MS analysis.

iST GFP-Trap Kit for mass spectrometry

Protein arrays (bead-based)

GST- & GFP- nanobodies for bead-based protein arrays e.g. Luminex®

Bead-based protein arrays using GFP-VHH and GST-VHH for capture of GFP- and GST-fusion proteins and their interacting partners.

GST- & GFP- nanobodies for bead-based protein arrays

Surface plasmon resonance (SPR)

Capture Surface for Biacore assays

GFP-VHH as capture surface reagent for direct immobilization of GFP-fusion proteins in BiacoreTM protein analysis assays.

Capture Surface for Biacore assays

Ubiquitination

Ubiquitination of GFP-tagged proteins

GFP-Trap for the analysis of candidate ubiquitinated proteins under harsh conditions.

Ubiquitination of GFP-tagged proteins