Application notes
Bio-Layer Interferometry (BLI)
Capturing COVID-19 antibodies for FortéBio BLI assays
Nano-CaptureLigands to immobilize different variants of the COVID-19 antibody CR3022 for determination of binding kinetics
Nano-CaptureLigands for COVID-19 antibodies capture in BLI binding kinetics assay
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 with RFP-tagged proteins
RFP-Trap for ChIP analysis in transiently expressing a mCherry fusion in mammalian cells.
Conjugation of fluorescent dyes to Nanobodies
Maleimide chemistry labeling protocol of Nanobodies
Recommendations for the site-directed labeling of ChromoTek Nanobodies containing 2 ectopic cysteines with thiol-reactive fluorescent dyes by maleimide chemistry.
NHS ester chemistry labeling protocol of Nanobodies
The protocol provides recommendations for the random labeling of ChromoTek Nanobodies containing surface-exposed lysines with NHS-reactive fluorescent dyes by NHS ester chemistry.
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.
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.
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.
Immunostaining
One-step incubation 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_Incubation_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..
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.
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.
Surface plasmon resonance (SPR)
Capturing GFP-fusion proteins for Biacore SPR assays
GFP-VHH as capture surface reagent for direct immobilization of GFP-fusion proteins in BiacoreTM protein analysis assays.
Ubiquitination
Ubiquitination of GFP-tagged proteins
GFP-Trap for the analysis of candidate ubiquitinated proteins under harsh conditions.