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HTRF mAb Anti-6His XL665-Conjugate, 1,000 Assay Points

XL665-labeled anti-6His antibody for capturing 6His-tagged proteins in protein/protein interaction assays.

For research use only. Not for use in diagnostic procedures.

View product information

Feature	Specification
Application	Protein-Protein Interaction

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XL665-labeled anti-6His antibody for capturing 6His-tagged proteins in protein/protein interaction assays.

For research use only. Not for use in diagnostic procedures.

View product information

Product Variants

Unit Size: 1,000 Assay Points

Part #:

61HISXLF

List Price

USD 262.25

Unit Size: 5,000 Assay Points

Part #:

61HISXLA

List Price

USD 609.47

Unit Size: 20,000 Assay Points

Part #:

61HISXLB

List Price

USD 1,911.00

Product information

Overview

MAb Anti-6HIS-XL665 is an IgG2a raised against a polyhistidine tagged fusion protein labeled with XL665. It recognizes synthetic polyhistidine or polyhistidine-tagged fusion protein.

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, receptor/ligand.

HTRF can detect a broad range of affinity constants ranging from picomolar to low millimolar.

Specifications

Other Specifications

Application	Protein-Protein Interaction
Brand	HTRF
Detection Modality	HTRF
Product Group	Fluorescent Reagent
Shipping Conditions	Shipped Ambient
Target Class	Binding Assay
Technology	TR-FRET
Therapeutic Area	Cardiovascular Infectious Diseases Inflammation Metabolism/Diabetes NASH/Fibrosis Neuroscience Oncology & Inflammation Rare Diseases
Unit Size	1,000 Assay Points

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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 signal is proportional to the binding of the 2 partners. In the example shown here: MAb Anti 6HIS-XL665 binds to the 6 His tagged partner A while partner B* binds to a specific Ab labeled with an HTRF acceptor. *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 acceptor for the detection.

Assay protocol

The example on the right describes the protocol using a 20 µL final assay volume for detecting an interaction between a 6 His-tagged partner A and a non-tagged partner B*. Dispense the 2 partners (10 µL), incubate, add MAb Anti 6HIS-XL665 (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.

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. Using 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.