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Encompassing Amino Acids: MLL 3745-end, WDR5 3745-end, other components 2-end
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 50 µl reaction mix (50 mM Tris, pH 8.8, 5 mM MgCl2, 4 M DTT, 20 µM 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.
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.
Description: 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.
Format: Aqueous buffer solution
Formulation: 45 mM Tris-HCl, pH 8.0, 124 mM NaCl, 2.4 mM KCl, 225 mM imidazole, 1 mM TCEP, 1 µM ZnCl2, and 10% glycerol
Genbank: NM_005933 (MLL1), NM_017588 (WDR5), NM_001105214 (Ash2L), NM_005057 (RbBP5), NM_032574 (DPY30)
Host Cell Line: E. coli
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal FLAG-tag (WDR5), N-terminal His-tag (others)
Uniprot: Q03164 (MLL1), P61964 (WDR5), Q9UBL3 (Ash2L), Q15291 (RbBP5), Q9C005 (DPY30)
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Patel, A., et al., J. Biol. Chem. 2009; 284(36): 24242-24256.
2. Mohan, M., et al., Nat. Rev. Cancer 2010; 10(10): 721-728.
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