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HTRF MAb Anti FLAG-XL665, 5,000 Assay Points

Picture anti-FLAG XL665
HTRF MAb Anti FLAG-XL665, 5,000 Assay Points
Picture anti-FLAG XL665
Picture anti-FLAG XL665
Picture anti-FLAG XL665
Picture anti-FLAG XL665
Picture anti-FLAG XL665

XL665-labeled anti-FLAG antibody for capturing FLAG-tagged proteins in protein/protein interaction assays.

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Feature Specification
Application Protein-Protein Interaction
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XL665-labeled anti-FLAG antibody for capturing FLAG-tagged proteins in protein/protein interaction assays.

View product information
Click to copy promo code to clipboard.
img-icon-10-off-white-yellow.svg

SAVE 10% on your first Revvity.com order. Use promo code below.

HELLO10

Terms and conditions apply.

Product Variants
Unit Size: 1,000 assay points
Part #:
61FGBXLF
List Price
USD 262.00
Your online price:
Unit Size: 5,000 assay points
Part #:
61FGBXLA
List Price
USD 609.00
Your online price:
Unit Size: 20,000 assay points
Part #:
61FGBXLB
List Price
USD 1,911.00
Your online price:
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
Total list price:
USD 609.00
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Picture anti-FLAG XL665
Picture anti-FLAG XL665
HTRF MAb Anti FLAG-XL665, 5,000 Assay Points
Picture anti-FLAG XL665
Picture anti-FLAG XL665
Picture anti-FLAG XL665
HTRF MAb Anti FLAG-XL665, 5,000 Assay Points
Picture anti-FLAG XL665
Picture anti-FLAG XL665

Product information

Overview

MAb Anti FLAG-XL665 is an antibody raised against FLAG® fusion proteins labeled with XL665. 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.

Specifications

Application
Protein-Protein Interaction
Brand
HTRF
Detection Modality
HTRF
Product Group
Fluorescent Reagent
Shipping Conditions
Shipped Ambient
Target
FLAG
Target Class
Binding Assay
Technology
TR-FRET
Unit Size
5,000 assay points

How it works

Principle of the HTRF Anti FLAG-XL665 assay

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-XL665 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 principle FLAG-XL665
Assay protocol of the HTRF Anti FLAG-XL665 assay

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 Anti FLAG-XL665 (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.

assay principle FLAG-XL665

Assay details

Build HTRF interaction assays for your specific application

Reagents are sold by the number of tests (20 µL reactions). Antibodies conjugated to XL665 or d2 are supplied on the basis of 20 ng of antibody per well. The amount of active moiety per vial is also provided (as well as the number of tracers per vial - see product description sheet). The active moiety is defined as the active part of a conjugate (e.g. antibody).

How do the number of tests relate to active moiety?

The average conjugate quantity per well reflects overall biological material content. Use of the active moiety amount is generally preferred to the quantity of total conjugate. For Cryptate and d2 conjugates, the total conjugate amount equals that of the active moiety, since the molecular weight of the label is negligible. This is not the case for XL665 labeled entities, for which the quantity of total conjugate will vary depending on the final molar ratio of the XL665 conjugate. However, the amount of active moiety provided by Revvity is constant and based on the number of tests ordered.

assay specifications active moiety
Recommended quantities of Cryptate and XL665 conjugates

Cryptate conjugates must not be excessive in order to prevent reader saturation and an unacceptable level of background. In most cases, a cryptate concentration of 1 to 5nM is appropriate, and will generate 20,000 to 80,000 cps at 620 nm depending on the HTRF compatible reader used. The XL665 conjugate must match its assay counterpart as closely as possible in order for the maximum number of biomolecules to be tagged with the XL665 acceptor. Thus, to detect a tagged molecule at an assay concentration of 20nM, the concentration of anti-Tag-XL665 should be equimolar or higher.

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