Revvity, ASBF-AIRF5-A10K, AlphaLISA SureFire Ultra Human IRF5 Biotin Free Aggregation Kit, 10,000 Assay Points

Overview
Interferon regulatory factor 5 (IRF5) is a transcription factor involved in innate immunity and inflammatory responses, regulating the expression of type I interferons (IFN-alpha and IFN-beta) and pro-inflammatory cytokines. Activated by pattern recognition receptors such as Toll-like receptors (TLR), MDA5, or STING in response to viral infection, IRF5 plays a crucial role in antiviral defense. In its inactive form, IRF5 resides in the cytoplasm of uninfected cells. Upon activation, it undergoes phosphorylation, dimerization, and nuclear localization to drive gene expression. Dysregulation of IRF5 is linked to autoimmune diseases such as lupus and rheumatoid arthritis, as well as inflammatory cancers, where it modulates tumor-associated macrophages.
The AlphaLISA SureFire Biotin-Free Human IRF5 Aggregate Detection Kit is a sandwich immunoassay for the quantitative detection of IRF5 aggregate 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 Biotin Free assay principle
The Aggregation-AlphaLISA SureFire Biotin Free assay uses a single antibody which recognizes an exposed epitope on the aggregated target protein, that is conjugated with either a CaptSure™ tag or with CaptSure3 tag. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure agent to specifically immobilize the antibody labeled with the CaptSure tag. Donor beads are coated with a proprietary CaptSure3 agent to capture the antibody labeled with the CaptSure3 tag. In the presence of aggregated target protein, the Donor and Acceptor beads are brought into close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing 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 Biotin Free 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 Biotin Free detection reagents. This protocol allows for 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 Biotin Free one-plate assay protocol
Detection of aggregated target protein with AlphaLISA SureFire Biotin Free reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining robust AlphaLISA SureFire Ultra quality.
Application-Cell Signaling
Automation Compatible-Yes
Brand-AlphaLISA SureFire Biotin-Free
Detection Modality-Alpha
Product Group-Kit
Protocol Time-2h at RT
Sample Volume-10 µL
Shipping Conditions-Shipped in Blue Ice
Target-IRF5
Target Class-Phosphoproteins
Target Species-Human
Technology-Alpha
Therapeutic Area-Inflammation
Unit Size-10,000 assay points