Skip to main content
Menu
US

HTRF CD16a (FcgRIIIa, V158) Cellular Binding Kit, 100 Assay Points

The CD16a cellular binding kit is designed for monitoring the binding of the IgG Fc region to the human CD16a receptor on live cells.

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).

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.
Feature Specification
Application Protein Quantification
Sample Volume 5 µL

The CD16a cellular binding kit is designed for monitoring the binding of the IgG Fc region to the human CD16a receptor on live cells.

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).

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: 100 Assay Points
Part #:
62C16PAG
List Price
USD 1,053.00
Unit Size: 500 Assay Points
Part #:
62C16PAE
List Price
USD 3,865.00
Unit Size: 10,000 Assay Points
Part #:
62C16PAF
List Price
USD 6,399.00

Overview

The CD16a (FcgRIIIa, V158) cellular binding assay is a simple and robust test to monitor binding of the IgG Fc region to the human CD16a receptor in live cells and to select the most potent antibody for use in Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC). The assay can also be used to assess the degree of antibody afucosylation. Two separate CD16a assays are available to specifically target the V158 or F158 variants. These assays, based on our innovative Tag-lite technology, can easily be automated to create a high-throughput solution for predicting ADCC.

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
5 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Biologics
Technology
TR-FRET
Therapeutic Area
Cardiovascular
Infectious Diseases
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Rare Diseases
Unit Size
100 Assay Points

Video gallery

How it works

Assay principle

The CD16a receptor cellular binding assay is a competitive assay involving HEK293 cells expressing a specific CD16a variant conjugated to Lumi4-terbium Cryptate, and human IgG1 labelled with d2. The unlabelled antibody or Fc tagged drug competes with the d2 labelled IgG1 for binding to the receptor, resulting in a decrease of FRET signal.

biomarkers-how-it-works-assay-principle-cd16a-62c16pae
Assay protocol

The CD16a receptor cellular binding assays (FcgIIIa V158 or F158) present only 1 incubation step after reagents distribution. Lumi4-terbium Cryptate pre-labelled cells are dispensed, followed by the addition of IgG1 labelled with d2 and the sample. After 2h incubation at RT, the signal can be read.

biomarkers-how-it-works-assay-protocol-cd16a-62c16pae

 

Assay validation

Specificity for IgG subclasses

Increasing concentrations of unlabeled IgGs were incubated in presence of a constant concentration of d2-labeled IgG1. The plate was then read on a PHERAstar microplate reader (BMG Labtech). As previously described, we confirm that the affinity of IgG1 for CD16a is better than that of IgG3 and considerably higher than IgG2 and IgG4 (Bruhns et al., Blood 2009).

assay-validation-cd16a-fcgriiia-v158-2
Afucosylation assessment

To evaluate the impact of IgG a-fucosylation on CD16a affinity, the same IgG with various degrees of fucosylation was tested following the previously described protocol. IgG affinity for CD16a and degree of a-fucosylation were very well correlated. These results confirm that a substantial gain of immunoglobulin affinity can be reached when the level of a-fucosylation is higher than 50%. The CD16a assay can therefore be used for the prediction of a-fucosylation degree and ADCC potential.

assay-validation-cd16a-fcgriiia-v158-2

 

Resources

Are you looking for resources, click on the resource type to explore further.

1-4 of 4 Resources
Application Note Icon
Application Note
High Throughput assays to compare the performance and stability of an engineered Fc silent antibody to a therapeutic antibody

Over the last decade, therapeutic antibodies have emerged as the predominant class of new drugs. Assays to functionally...

Brochure Icon
Brochure
HTRF assays and reagents catalog

Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...

Guide Icon
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

Flyer Icon
Flyer
Therapeutic development with robust cellular binding assays.

From the chemistry of our assays to the collaborative nature of our relationship with customers, Revvity is founded on the belief...

Scroll Icon