Our multiplex immunoassays use the FirePlex® particle technology to enable the quantitative measurement of many human proteins and peptides at the same time using as little as 12.5 μL of sample per well. Assay run-time is 3.5 hours, followed by particle analysis using a validated flow cytometer model and data analysis using our integrated, free-of-charge FirePlex Analysis Workbench software.
FirePlex immunoassays offer: - Measurement of multiple analytes in the same well, thus conserving time and precious samples. - Flexibility to select either from our catalogue of pre-designed panels or build custom panels from our large antibody pairs portfolio. - Antibody pairs that are validated across a broad set of biological sample types, and provide sensitive and reproducible quantitation of analytes in a given sample.
Important - Use of the FirePlex Human Inflammation - Immunoassay Panel requires the purchase of: - a) A FirePlex Immunoassay Human Core Reagent Kit (ab208203); and - b) Human Immunoassay Protein Standard Mix A (ab227934).
FirePlex® is a registered trade mark in the United States and is registered as a European Union Trade Mark.
The following information provides the sensitivity, range, CVs, required Protein Standard Mix and links to the individual target kit, where more information regarding recommeneded dilutions can be found.
TNF alpha: Secreted and Cell membrane.
Interferon gamma: Secreted.
IL1 beta: Cytoplasm, cytosol. Lysosome. Secreted, exosome. Cytoplasmic vesicle, autophagosome. Secreted. The precursor is cytosolic. In response to inflammasome-activating signals, such as ATP for NLRP3 inflammasome or bacterial flagellin for NLRC4 inflammasome, cleaved and secreted. IL1B lacks any known signal sequence and the pathway(s) of its secretion is(are) not yet fully understood (PubMed:24201029). On the basis of experimental results, several unconventional secretion mechanisms have been proposed. 1. Secretion via secretory lysosomes: a fraction of CASP1 and IL1B precursor may be incorporated, by a yet undefined mechanism, into secretory lysosomes that undergo Ca(2+)-dependent exocytosis with release of mature IL1B (PubMed:15192144). 2. Secretory autophagy: IL1B-containing autophagosomes may fuse with endosomes or multivesicular bodies (MVBs) and then merge with the plasma membrane releasing soluble IL1B or IL1B-containing exosomes (PubMed:24201029). However, autophagy impacts IL1B production at several levels and its role in secretion is still controversial. 3. Secretion via exosomes: ATP-activation of P2RX7 leads to the formation of MVBs containing exosomes with entrapped IL1B, CASP1 and other inflammasome components. These MVBs undergo exocytosis with the release of exosomes. The release of soluble IL1B occurs after the lysis of exosome membranes (By similarity). 4. Secretion by microvesicle shedding: activation of the ATP receptor P2RX7 may induce an immediate shedding of membrane-derived microvesicles containing IL1B and possibly inflammasome components. The cytokine is then released in the extracellular compartment after microvesicle lysis (PubMed:11728343). 5. Release by translocation through permeabilized plasma membrane. This may occur in cells undergoing pyroptosis due to sustained activation of the inflammasome (By similarity). These mechanisms may not be not mutually exclusive.