MAb Anti 6HIS-Tb cryptate HTRF®

Tb cryptate-labeled anti-6His antibody for capturing 6His-tagged proteins in protein/protein interaction assays.
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  • High affinity High affinity
Tb cryptate-labeled anti-6His antibody for capturing 6His-tagged proteins in protein/protein interaction assays.


MAb Anti-6HIS-Tb cryptate is an IgG2a raised against a polyhistidine tagged fusion protein labeled with Tb. 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.



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-Tb cryptate 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 principle of detection assay using HTRF MAb Anti 6HIS-Tb cryptate

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-Tb cryptate (5 µL) and anti-partner B labeled with acceptor. (5 µL), incubate and read. *partner B can also be biotinylated, tagged, Fc fused or directly labeled. In these cases use for the detection, the corresponding HTRF reagent (anti-Tag, anti species, protA, Streptavidin), labeled with acceptor..
assay protocol of detection assay using HTRF MAb Anti 6HIS-Tb cryptate

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 Cisbio, is constant and based on the number of tests ordered.
description of the active moiety in an HTRF conjugate

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.

Applications of HTRF and Tag-lite-lite Assays for HTP Antibody Screening

In collaboration with Bristol Myers Squibb - Scientific Presentations

Academia/Industry Cross-Fertilization through Translational Research

In collaboration with Scripps - Scientific Presentations

HTRF-powered detection of ubiquitin-like protein (UBL) signaling and interactions

In collaboration with Sanford/Burnham - Scientific Presentations

The use of HTRF in biologics discovery

In collaboration with MedImmune - Scientific Presentations

HTRF: One technology, many uses in a pre-clinical laboratory setting

Benefits and considerations of HTRF - Scientific Presentations

Exploiting HTRF for novel drug classes: Stabilizing 14-3-3 protein-protein interactions

In collaboration with Lead Discovery Center GmbH - Scientific Presentations

HTRF a versatil approach for 7TM drug discovery

In collaboration with MRC - Scientific Presentations

HTRF protein-protein interaction reagents

Benefit from unlimited flexibility for your assay development - Flyers

Addressing the interactome with protein-protein assays

Cover all PPIs with one approach - Brochures

HTRF -A Beneficial Tool for Lead Optimization of Biotherapeutics

In collaboration with Boehringer Ingelheim - Posters

Product Insert Anti 6HIS-Tb / 61HISTLF-61HISTLA-61HISTLB

61HISTLF-61HISTLA-61HISTLB - Product Insert

HTRF PPI your dream assay served on a plate

Sandwiches aren't just for eating - Infographics

Best practices for pharmacological characterization of PPI inhibitors

Easy pharmacological characterization of PPI modulators. - Technical Notes

HTRF assays handle low- to high affinity protein-protein interactions

Deciphering low- and high affinity interactions - Application Notes

Nuclear receptor ligand identification with HTRF

Monitoring nuclear receptor binding with HTRF assays - Application Notes

HTRF addresses large protein-protein interaction complexes

Challenge large complexes with HTRF assays - Application Notes

A brief history of Protein-Protein Interactions

How well do you know PPI? - Infographics

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Virology research solutions using HTRF Protein-Protein Interaction assays

See how peer researchers challenge the viral life cycle with PPI assays - Application Notes

Solutions for CAR-T research

Advance your CAR-T cell research - Flyers

Safety Data Sheet Anti 6HIS-Tb / 61HISTLA

61HISTLA - Safety Data Sheet

Batch Information Anti 6HIS-Tb / 20250105

61HISTLA Batch 13A - Batch Information

Safety Data Sheet Anti 6HIS-Tb / 61HISTLB

61HISTLB - Safety Data Sheet

Batch Information Anti 6HIS-Tb / 20250105

61HISTLB Batch 13A - Batch Information

Batch Information Anti 6HIS-Tb / 20240415

61HISTLF Batch 11B - Batch Information

Batch Information Anti 6HIS-Tb / 20250105

61HISTLF Batch 13A - Batch Information

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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