AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
IL1α and IL1b are central players of the immune response, displaying roles in inflammation both at local and systemic levels. Despite they seem to display very similar functions, these proteins are encoded by two independent genes sharing only ~30% identity. IL1b is synthesized as a 31 kDa precursor that is cleaved by Caspase-1 (ICE) into the active 17 kDa form, and eventually released into the extracellular space. Its production has been reported in many cell types including brain and, importantly, monocytic and periferal blood mononuclear cells. After binding to its receptor, IL-1RI, IL1b triggers a cascade of kinase signaling pathways that lead to the activation of transcription factors like NF-kB and AP-1, eventually activating the expression of genes such as MIP-2 and C-reactive protein.
This kit is designed for the quantitative determination of human IL1b in serum, buffered solution or cell culture medium using a homogeneous AlphaLISA assay (no wash steps).
In AlphaLISA immunoassay buffer:
- Lower detection limit (LDL): 0.65 pg/mL
- Dynamic range: 0.65-30,000 pg/mL
Sensitivity can be increased by increasing the volume of analyte in the assay.
In analyte-depleted serum:
- Lower detection limit (LDL): 4.0 pg/mL
- Dynamic range: 4.0-30,000 pg/mL
For research use only.
The data was generated using a white OptiPlate-384 microplate and an EnVision-Alpha Reader 2102.
Each Kit Contains:
- AlphaLISA Acceptor beads coated with Anti-Analyte Antibody #1
- Streptavidin-coated Donor beads
- Biotinylated Anti-Analyte Antibody #2
- Lyophilized analyte
- AlphaLISA Immunoassay Buffer (10X).
Buffer and lyophilized analyte can be ordered separately.