Product Description
Overview
MAb Anti FLAG-d2 is an antibody raised against FLAG® fusion proteins labeled with d2. The anti-FLAG antibody will recognize the FLAG sequence at the N-terminus, intra, or C-terminus of FLAG fusion proteins.
This reagent can be used in both biochemical and cellular formats to study a wide variety of interactions: protein/protein, protein/peptide, protein/DNA, protein/RNA, protein/carbohydrate, protein/small molecule, and receptor/ligand.
HTRF can detect a broad range of affinity constants ranging from picomolar to low millimolar.
HTRF assays offer many advantages over other technologies:
Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
Assay principle
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 resulting signal is proportional to the binding of the 2 partners. In the example shown here, MAb Anti FLAG-d2 binds to the FLAG tagged partner A, while partner B* binds to a specific Ab labeled with an HTRF donor.
*partner B can also be biotinylated, tagged, or Fc fused. In these cases, use the corresponding HTRF reagent (anti-Tag, anti-species, protA, Streptavidin) labeled with a donor for the detection.
Assay protocol
The example on the right describes the protocol using a 20 µL final assay volume for the detection of an interaction between a FLAG-tagged partner A and a non-tagged partner B*. Dispense the 2 partners (10 µL), incubate, add MAb Anti FLAG-d2 (5 µL) and anti-partner B labeled with donor (5 µL), then 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 a donor for the detection. Application-Protein-Protein Interaction
Brand-HTRF
Detection Modality-HTRF
Product Group-Fluorescent Reagent
Shipping Conditions-Shipped Ambient
Target Class-Binding Assay
Technology-TR-FRET
Unit Size-1,000 assay points
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Collaboration
Tony Tang
Email: Tony.Tang@iright.com
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