MLL4 (KMT2B): WDR5 Binding Chemiluminescent Assay Kit
The MLL4 (KMT2B): WDR5 Binding Chemiluminescent Assay Kit is an ELISA-based assay designed to measure the binding between MLL4 (mixed lineage leukemia protein-4, also known as KMT2B) and WDR5 (WD40 repeat-containing protein 5) for screening and profiling applications. The MLL4 (KMT2B): WDR5 Binding Chemiluminescent Assay comes with enough purified MLL4 (amino acids 2490-2715) and WDR5 proteins, primary and secondary antibody, assay buffer, and detection reagent for 100 enzyme reactions.
Figure 1. MLL4 (KMT2B): WDR5 Binding Chemiluminescent Assay Kit schematic.
A 96-well plate is coated with MLL4 protein. After coating and blocking, WDR5 is added in an optimized assay buffer. Next, unbound WDR5 is washed away, and the plate is incubated with a primary antibody followed by a secondary HRP-conjugated antibody. 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 WDR5 binding to MLL4 (KMT2B).
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- 1x TBS (Tris Buffer Saline) Buffer
- TBST Buffer (1x TBS, containing 0.05% Tween-20)
- Luminometer or microplate reader capable of reading chemiluminescence
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| 11344 | MLL4, His-Tag* | 2.5 µg | -80°C |
| 79187 | WDR5, FLAG-Tag* | 2.5 µg | -80°C |
| 3x PL-02 Assay Buffer | 50 µl | -80°C | |
| Primary Antibody AB31 | 6 µl | -80°C | |
| 52130H | Secondary HRP-Labeled Antibody 1 | 10 µl | -80°C |
| Blocking Buffer 8 | 50 ml | +4°C | |
| 79670 | ELISA ECL Substrate A (translucent bottle) | 6 ml | Room Temp |
| ELISA ECL Substrate B (brown bottle) | 6 ml | Room Temp | |
| 79837 | 96-well module plate | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
The Mixed Lineage Leukemia-4 (MLL4, also known as KMT2B) protein belongs to the SET1/MLL family which consists of six (MLL1-4/KMT2A-2D, SET1A/KMT2F, and SET1B/KMT2G) major methyltransferases in mammals. MLL4 is a histone-H3 lysine-4 (H3K4) methyltransferase that promotes H3K4 mono-/di-/tri-methylation, a conserved trait of euchromatin associated with transcriptional activation. MLL4 is a critical player in memory formation. MLL4 forms a complex with RbBP5 (retinoblastoma-binding protein 5), ASH2L (Absent, small, homeotic disks-2-like), WDR5 (WD40 repeat-containing protein 5), and DPY30 (DumPY protein 30) to catalyze methylation of H3K4. WDR5 represents a therapeutically exploitable target for cancer treatment as it plays a crucial role in MLL1 complex assembly and disassembly. WDR5 has two protein interaction sites: the WDR5-interacting (WIN) binding site and the WDR5-binding-motif (WBM) site. MLL1-4 forms the complex via WIN binding site, while RbBP5 is bound to WBM site, which is also the site for MYC oncoproteins interaction. The addition of inhibitors that competitively bind to WIN or WBM sites has been shown to disrupt MLL activity as well as to displace MYC from chromatin and therefore disabling its tumorigenic function.
Wang L., et al., 2021 Biochem Soc Trans 49(6):1041-1054.
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