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Application
The Monoclonal Anti-His Tag Antibody, Human IgG1 FA-Labeled is a universal tool for TR-FRET assays that can bind His-tagged proteins. This product uses specific anti-His antibody labeled with fluorescent dyes and it can be used in combination with other Donors covalently conjugated with Eu³⁺ chelate. When the Donor and Acceptor come into close proximity (within a distance of less than 10 nm), a FRET reaction occurs: the 620 nm signal emitted by the Donor upon excitation by a specific light source is received by the Acceptor, which then emits a 665 nm signal.Product Specifications
SourceThis Monoclonal Anti-His Tag Antibody, Human IgG1 is a chimeric monoclonal antibody recombinantly expressed from HEK293, which combines the variable region of a mouse monoclonal antibody with Human constant domain.SpecificityThis antibody specifically reacts with His tag proteins, and can recognizes C-terminal, N-terminal, and internal His tagged fusion proteins.FormulationLyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.Physical AppearancePowderStorage (unopened)-20°C to -70°CStorage (opened)-70°CUnit Size1000 tests & 5000 tests & 20000 testsTechnologyTR-FRETIntended UseFor binding the his-tagged protein as acceptor in TR-FRET AssayReconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
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Bioactivity-TR-FRET
Please refer to DS document for the assay protocol.
In this TR-FRET assay, Streptavidin Protein Europium chelate (Cat. No. FRT-TD09) is used as the Donor and Monoclonal Anti-His Tag Antibody, Human IgG1 FA-Labeled (Cat. No. FRT-TA04) is used as the Acceptor. Biotinylated Human PD-1, Fc,Avitag (Cat. No. PD1-H82F1), at 12.5 ng/well, can bind to the Human PD-L1, His Tag (Cat. No. PD1-H5229) with a linear detection range of 10-0.02 μg/mL (QC tested).
In this TR-FRET assay, Streptavidin Protein Europium chelate (Cat. No. FRT-TD09) is used as the Donor and Monoclonal Anti-His Tag Antibody, Human IgG1 FA-Labeled (Cat. No. FRT-TA04) is used as the Acceptor. An inhibition Assay was performed to evaluate the interaction between Biotinylated Human PD-1, Fc, Avi tag (Cat. No. PD1-H82F1) and Human PD-L1, His Tag (Cat. No. PD1-H5229) using Monoclonal Anti-Human PD-1 Antibody, Human IgG4 resulting in a typical IC50 of 2.230 μg/mL (Routinely tested).
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