MLL1/WARD complex of human MLL1 (GenBank Accession No. NM_005933,) a.a. 3745-end with N-terminal His-tag and MW= 27 kDa, human WDR5 (GenBank Accession No. NM_017588), a.a. 22-end with N-terminal FLAG-tag and MW= 35 kDa, human Ash2L (GenBank Accession No. NM_001105214), a.a. 2-end with N-terminal His-tag and MW= 61 kDa, human RbBP5 (GenBank Accession No. NM_005057), a.a. 2-end with N-terminal His-tag and MW= 60 kDa, and human DPY30 (GenBank Accession No. NM_032574) a.a. 2-end with N-terminal His-tag and MW=12 kDa individually expressed in an E. coli expression system.
Host cell line: E. coli
Background
MLL is a histone methyltransferase that plays an essential role in early development and hematopoiesis. Part of a multiprotein complex that mediates both methylation of Lys4 of histone H3 (H3K4me) complex and acetylation of Lys16 of histone H4 (H4K16ac). Has weak methyltransferase activity by itself, and requires other component of the MLL1/MLL complex to obtain full methyltransferase activity. Multiple chromosomal translocations involving this gene are the cause of certain acute lymphoid leukemias and acute myeloid leukemias.
Immunogen Region
MLL 3745-end, WDR5 3745-end, other components 2-end
45 mM Tris-HCl, pH 8.0, 124 mM NaCl, 2.4 mM KCl, 225 mM imidazole, 1 mM TCEP, 1 uM ZnCl2, and 10% glycerol
Species
Human
Application
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling
Notes
50 ul reaction mix (50 mM Tris, pH 8.8, 5 mM MgCl2, 4 M DTT, 20 uM S-adenosylmethionine, and 5-50 ng MLL complex) add to the wells coated with the substrate. Incubate at room temperature for 1 hr. Add antibody against methylated K4 residue of histone H3, incubate 1 hr. Then, incubate 30 minutes with the secondary HRP-labeled antibody. Finally, add HRP chemiluminescent substrate and read luminescence.