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Application: Study and screen compounds that inhibit the binding of c-Myc to WDR5 for drug discovery in high throughput screening (HTS) applications.
Background: WDR5 (WD40 repeat-containing protein 5) is a protein that is part of the histone methyltransferase complex SET1/MLL (Protein Mixed Lineage Leukemia) which catalyzes histone 3 lysine 4 methylation (H3K4me). In addition to its direct role in gene regulation by impacting histone methylation, WDR5 can induce transcription of c-Myc induced genes, by bringing c-Myc to the appropriate regions of the chromosome. WDR5 has two protein interaction sites: the WDR5-interacting (WIN) binding site and the WDR5-binding-motif (WBM) site. MLL1 forms the complex via the WIN binding site, while RbBP5 is bound to WBM site, which is also the site for c-Myc oncoproteins interaction. WDR5 is found at high levels in certain cancers, as is c-Myc. The addition of inhibitors that competitively bind to WIN or WBM sites has been shown to disrupt MLL activity as well as displace c-Myc from chromatin and therefore disabling its tumorigenic function.
Contraindications: This assay kit is compatible with up to 1% final DMSO concentration.
Description: The c-Myc: WDR5 [Biotinylated] Binding Chemiluminescent Assay Kit is an ELISA designed to measure the binding between c-Myc and WDR5 (WD40 repeat domain 5) protein for screening and profiling applications. This kit comes with enough purified c-Myc (amino acids 2-454(end)) and biotinylated WDR5 (amino acids 2-334(end)), MLL1 (amino acids 3745-end), streptavidin-HRP, assay buffer, and detection reagent for 100 enzyme reactions.Figure 1. c-Myc: WDR5 [Biotinylated] Binding Chemiluminescent Assay Kit schematic. A 96-well plate is coated with c-Myc protein. After coating and blocking, MLL1 and biotinylated WDR5 are added in an optimized assay buffer. Next, unbound biotinylated WDR5-MLL complexes are washed away, and the plate is incubated with streptavidin-HRP. After a final wash, 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 WDR5 binding to c-Myc.Interested in having us perform the screening and profiling of inhibitors or blocking antibodies? Visit our Methyltransferase Screening& Profiling Services
Format: Catalog #NameAmount Storage 40453c-Myc, His-tag*5 µg-80°C80016WDR5, His-Tag, Biotin-Labeled * 5 µg-80°C79186MLL1, His-Tag*5 µg-80°C82952PP-03 Buffer10 ml-20°C79728Blocking Buffer 225 ml+4°C79742Streptavidin HRP10 µl+4°C79670ELISA ECL Substrate A (translucent bottle)6 mlRoom TempELISA ECL Substrate B (brown bottle)6 mlRoom Temp79699White 96-well microplate1Room Temp*The concentration of the protein is lot-specific and will be indicated on the tube.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: This kit comes in a convenient 96-well format with enough purified c-Myc, biotinylated WDR5, MLL1, streptavidin-HRP, assay buffer, and detection reagent for 100 enzyme reactions.
Target: C-MYC, WDR5
Uniprot: c-Myc: P01106, WDR5: P61964
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Thomas L., et al., 2020 Molecular and Cellular Oncology 7(2): 1709388.
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