Revvity, 64AAV1PEH, HTRF AAV1 Detection Kit, 10,000 Assay Points

Overview
Adeno-associated virus (AA) vectors are the leading platform for gene delivery for the treatment of a variety of human diseases. AAV Serotype 1 (AAV1) is an efficient vector for gene delivery to skeletal muscle or the central nervous system. The AAV1 kit is designed to detect and quantify AAV1 particles in an easy-to-use, no-wash format. The simple and robust procedure benefits from increased throughput compared to ELISA.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
AAV1 Capsid assay principle
The Adeno-Associated Virus serotype 1 (AAV1) assay measures AAV1 capsid in cell supernatant or cell lysate. The assay uses two anti-AAV1 antibodies: one coupled to HRP and which binds to anti-HRP d2 (acceptor) in premix 1, and the other coupled to biotin and which binds to Streptavidin Eu-cryptate (donor) in premix 2. In presence of AAV1 capsid in a cell extract or supernatant, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the capsid present in the sample, and provides a means of assessing any changes caused by experimental variability under a no-wash assay format.

 

AAV1 Capsid assay protocol
The AAV1 Capsid assay protocol using a 384-well small volume white plate is described on the right. 5 µL of sample or standard and 5 µL of diluent are dispensed directly into the plate for detection by HTRF® reagents. The Biotin antibody anti-AAV1 is pre-mixed with Streptavidin labeled with the donor, and HRP anti-AAV1 was pre-mixed with anti-HRP labeled with the acceptor. 5 µL of each premix were added. The assay can be run in up to a 1536-well format by simply resizing each addition volume proportionally. Application-Bioprocessing
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Product Group-Kit
Sample Volume-5 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Viral Particles
Technology-TR-FRET
Unit Size-10,000 assay points