Pages

Pretest TriCEPS Service and Kit

Flow TriCEPS™ Service and Kit


lrc-pretest-triceps-kit-2016-dualsystems

TriCEPS V2.0 mediated selection of the cells for target identification using Flow Cytometry (FACS).


Now, Dualsystems provides TriCEPS V2.0 for choosing the best cell type for your LRC-TriCEPS experiment to identify the targets of your ligand (peptide, protein, Antibody, ADC or other primary amine containing molecules). TriCEPS V2.0 couples to your ligand at the primary amines (i.e. Lysines and N-terminus) and since TriCEPS contains a biotin it can be detected using Flow Cytometry by attaching a fluorophore.

TriCEPS V2.0 is a tool to support the selection of the right cells to identify the unknown target receptors. Further, it can be used as Flow Cytometric reagent for testing binding of proteins or other primary amines containing molecules to the cell surface.

Pretests are provided as service by Dualsystems Biotech AG or pretest kits can be purchased to perform the pretests in house.

TriCEPS V.2.0 based pretests



Figure 1: Flow cytometry measurements of MDA-MB231 cells. TriCEPS coupled glycine is used as a negative control. TriCEPS coupled transferrin is positive control. Dashed line indicates non-treated cells. Streptavidin-Phycoerythrin (PE) is used as a second step reagent for indirect immunofluorescent straining of biotinylated TriCEPS V2.0


Cellular responses to ligands such as peptides, proteins, pharmaceutical drugs or entire pathogens are generally mediated through interactions with proteins expressed at the cell surface. The LRC-TriCEPS kit is designed to directly identify the target proteins of your ligand on the living cells using near-physiological conditions.

Dualsystems offers now the pretest TriCEPS experiments as services or as kit.

The pretest is designed to be used to screen different cell types to decide which cells to use for the identification of the unknown targets of your ligand of interest later in the LRC-TriCEPS main experiment.

After successful completion of all pretests the main experiment can be planned and carried out as described in the LRC-TriCEPS user manual choosing one of the service options provided by Dualsystems Biotech AG.

Merken


Flow-TriCEPS Flyer


Further, in case functional assays are available TriCEPS coupled ligands can be tested to see if a similar output can be achieved with a TriCEPS coupled ligand compared to a ligand that is not coupled to TriCEPS.

In addition, TriCEPS V2.0 can be utilized as Flow Cytometry (FACS) reagent for detecting binding of primary amine containing molecules to the cells.

In a first step, the TriCEPS v.2.0 molecule is coupled to the ligand of interest (peptide, protein, Antibody, ADC or other primary amine containing molecules) and to the positive control ligand (e.g. transferrin) and negative control ligand (e.g. glycine). This coupling reaction is tested with Dot blot to assess if the coupling worked.

Then, the TriCEPS coupled ligands are added to the non-oxidized cells to show that the ligand binds to the unknown targets at the cell surface and TriCEPS does not interfere with the ligand receptor interaction.

Further, Flow Cytometry tests with TriCEPS coupled ligand on oxidized cells can be performed to test cell viability and ligand target binding as it will occur in the main LRC-TriCEPS experiment.

After successful completion of all pretests the main experiment can be planned and carried out as described in the LRC-TriCEPS user manual choosing one of the service options provided by Dualsystems Biotech AG.