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HTRF mAb Anti-HA Eu-Conjugate, 5,000 Assay Points

Eu cryptate-labeled anti-HA antibody for capturing HA-tagged proteins in protein/protein interaction assays.

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

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Feature Specification
Application Protein-Protein Interaction

Eu cryptate-labeled anti-HA antibody for capturing HA-tagged proteins in protein/protein interaction assays.

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

Click to copy promo code to clipboard.
SAVE 15% NOW on online orders. Use promo code below.
YEAREND15
Offer valid until 12/15. Terms and conditions apply.
Product Variants
Unit Size: 5,000 Assay Points
Part #:
610HAKLA
List Price
USD 609.47
Unit Size: 20,000 Assay Points
Part #:
610HAKLB
List Price
USD 1,911.00

Overview

MAb Anti HA-Eu cryptate is an IgG2b raised against the twelve-amino acid peptide sequence CYPYDVPDYASL labeled with Eu. It recognizes the influenza virus hemagglutinin epitope (YPYDVPDYA) which has been used extensively as a general epitope tag in expression vectors.

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, 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 Class
Binding Assay
Technology
TR-FRET
Therapeutic Area
Cardiovascular
Infectious Diseases
Inflammation
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
Rare Diseases
Unit Size
5,000 Assay Points

Video gallery

Citations

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 HA-Eu cryptate binds to the HA 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.

htrf mab anti ha-eu cryptate
Assay protocol

The example on the right describes the protocol using a 20 µL final assay volume for detecting an interaction between a HA-tagged partner A and a non-tagged partner B*. Dispense the 2 partners (10 µL), incubate, add MAb Anti HA-Eu 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 the corresponding HTRF reagent (anti-Tag, anti species, protA, Streptavidin) labeled with acceptor for the detection.

htrf mab anti ha-eu cryptate

Assay details

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.

Active moiety in 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.

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