Revvity, ALSU-AIRF5-A-HV, AlphaLISA Human IRF5 Aggregate Detection Kit, 100 Assay Points

Overview
Interferon regulator factor 5, IRF5, is a transcriptional regulator of type I interferon (IFN-alpha and IFN-beta)-dependent immune responses. This transcription factor plays a key role in the innate immune response against DNA and RNA viruses. IRF5 is present in the cytoplasm of uninfected cells in an inactive form. Various mediators of the pathways that follow viral infection can overexpress and phosphorylate IRF5. IRF5 is associated with various cancers and autoimmune diseases.
The AlphaLISA SureFire Ultra Human IRF5 Aggregate Detection Kit is a sandwich immunoassay for the quantitative detection of aggregate IRF5 in cellular lysates, using Alpha Technology.
Formats:

The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.
The 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.

AlphaLISA SureFire Ultra kits are compatible with:

Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies

AlphaLISA SureFire Ultra kits can be used for:

Cellular kinase assays
Receptor activation studies
High-throughput screening for preclinical studies
How it works
Aggregation-AlphaLISA SureFire Ultra assay principle
The Aggregation-AlphaLISA SureFire Ultra assay measures the aggregates level of a protein target in a cell lysate.
The Aggregation-AlphaLISA Surefire Ultra assay uses one antibody which recognizes an exposed epitope on the aggregated target protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure tag. Donor beads are coated with streptavidin to capture the detection antibody, which is biotinylated. In the presence of targeted protein, the antibody brings the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, enabling the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of aggregated protein present in the sample.

 

Aggregation-AlphaLISA SureFire Ultra two-plate assay protocol
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well Optiplate plate before the addition of Aggregation-AlphaLISA SureFire Ultra detection reagents. This protocol enables the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.

Aggregation-AlphaLISA SureFire Ultra one-plate assay protocol
Detection of aggregated target protein with AlphaLISA SureFire Ultra reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust AlphaLISA SureFire Ultra quality.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Ultra
Detection Modality-Alpha
Product Group-Kit
Protocol Time-2h at RT
Shipping Conditions-Shipped in Blue Ice
Target-IRF5
Target Class-Phosphoproteins
Target Species-Human
Technology-Alpha
Therapeutic Area-Autoimmunity Oncology
Unit Size-100 assay points