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Application: Screen small molecule inhibitors that block TSLP binding to TSLPR.
Background: TSLP (thymic stromal lymphopoietin) is a type I cytokine that functions as an alarmin and growth factor in the immune system. It is involved in type 2 immune responses, TH2 (T helper 2 cells) responses, and the maturation and recruitment of dendritic cells (DCs), T cells, B cells, neutrophils, mast cells, and other lymphoid cells. It can be produced by epithelial and stromal cells in the lung, skin, and gastric systems, but also by DCs, basophils and mast cells. Its expression can be induced by infections, pro-inflammatory cytokines, proteases, and even mechanical injury. For instance, it can be produced in the lungs in response to infection with influenza or rhinovirus. Its role as an alarmin can result in increasing inflammation. TSLP is linked to allergic reactions such as asthma, atopic dermatitis, and food allergies, by inducing the expression of OX40L, CD80 and CD86 and stimulating CD4+ T cells. In 2021, the TLSP-neutralizing antibody tezepelumab was approved for the treatment of severe asthma. Targeting TSLP is an active area of investigation with ongoing clinical trials for the treatment of autoimmune disorders.
Contraindications: The DMSO concentration in the final reaction should be ≤3%.
Description: The TSLPR:TSLP [Biotinylated] Small Molecule Inhibitor Screening Chemiluminescence Assay Kit is designed for screening and profiling small molecules that block the binding between TSLPR (thymic stromal lymphopoietin receptor) and TSLP. This kit comes in a convenient 96-well format, with enough recombinant human biotin-labeled TSLP (amino acids 29-159), human TSLPR (amino acids 25-231), streptavidin-HRP, and assay buffer for 100 reactions.A 96-well plate is coated with TSLPR protein. After coating and blocking, TSLP is added in an optimized assay buffer. Unbound TSLP is washed away, and the plate is incubated with streptavidin-HRP. Finally, ELISA ECL substrate is added to produce chemiluminescence that can be measured using a chemiluminescence reader. The chemiluminescence signal is proportional to the efficacy of TSLP binding to TSLPR.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: TSLPR: Q9HC73; TSLP: Q969D9
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)