Development of an HTRF method to determine the stoichiometry of subunits...
MAb Anti cmyc-d2
- High affinity
MAb Anti cmyc-d2 is an IgG1 raised against a synthetic peptide corresponding to the 408-439 sequence of human c-myc protein labeled with d2. It recognizes the EQKLISEEDL motif and is specific for human c-myc, although a slight cross-reaction has been observed with murine c-myc at high antibody concentration.
This reagent can be used in both biochemical and cellular formats to study a wide variety of interactions: protein/protein, protein/peptide, protein/DNA, receptor/ligand..
HTRF can detect a broad range of affinity constants ranging from picomolar to low millimolar.
- LARGE COMPLEXES DETECTION
- BATCH-TO-BATCH REPRODUCIBILITY
- LOW TO HIGH KD DETECTION
In an HTRF interaction assay, one partner is labeled (directly or indirectly) with the donor, and the other with the acceptor (again, directly or indirectly). The intensity of the signal is proportional to the binding of the 2 partners.
In the example shown here: MAb Anti cmyc-d2 binds to the c-Myc tagged partner A while partner B* binds to a specific Ab labeled with an HTRF donor.
*partner B can also be biotinylated, tagged, Fc fused. In these cases, use the corresponding HTRF reagent (anti-Tag, anti-species, protA, Streptavidin) labeled with donor for the detection.
The example on the right describes the protocol using a 20 µL final assay volume for detecting an interaction between a c-Myc-tagged partner A and a non-tagged partner B*.
Dispense the 2 partners (10 µL), incubate, add MAb Anti cmyc-d2 (5 µL) and anti-partner B labeled with donor (5 µL), incubate and read.
*partner B can also be biotinylated, tagged, Fc fused or directly labeled. In these cases, use the corresponding HTRF reagent (anti-Tag, anti species, protA, Streptavidin) labeled with donor for the detection.
61MYCDAF-61MYCDAA-61MYCDAB - Product Insert
Easy pharmacological characterization of PPI modulators. - Technical Notes
Deciphering low- and high affinity interactions - Application Notes
Monitoring nuclear receptor binding with HTRF assays - Application Notes
Challenge large complexes with HTRF assays - Application Notes
Get the brochure about technology comparison. - Brochures
See how peer researchers challenge the viral life cycle with PPI assays - Application Notes