Histone methyltransferase that catalyzes methyl group transfer from S-adenosyl-L-methionine to the epsilon-amino group of 'Lys-4' of histone H3 (H3K4) via a non-processive mechanism. Part of chromatin remodeling machinery, forms H3K4me1, H3K4me2 and H3K4me3 methylation marks at active chromatin sites where transcription and DNA repair take place. Responsible for H3K4me3 enriched promoters and transcriptional programming of inner mass stem cells and neuron progenitors during embryogenesis. Required for H3K4me1 mark at stalled replication forks. Mediates FANCD2-dependent nucleosome remodeling and RAD51 nucleofilaments stabilization at reversed forks, protecting them from nucleolytic degradation. Does not methylate 'Lys-4' of histone H3 if the neighboring 'Lys-9' residue is already methylated.
Background References
1. Kummeling J., et al. Characterization of SETD1A haploinsufficiency in humans and Drosophila defines a novel neurodevelopmental syndrome. Mol. Psychiatry 26:2013-2024 (2021)
2. Wysocka J., Myers M.P., Laherty C.D., Eisenman R.N., Herr W. Human Sin3 deacetylase and trithorax-related Set1/Ash2 histone H3-K4 methyltransferase are tethered together selectively by the cell- proliferation factor HCF-1. Genes Dev. 17:896-911 (2003)
Flow cytometric analysis of 293T cells labeling hSET1/SET1.
Cells were fixed and permeabilized.Then stained with the primary antibody (HA721226, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Flow cytometric analysis of HepG2 cells labeling hSET1/SET1.
Cells were fixed and permeabilized.Then stained with the primary antibody (HA721226, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of hSET1/SET1 on different lysates with Rabbit anti-hSET1/SET1 antibody (HA721226) at 1/1,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 2: HEK 293T cell lysate Lane 3: Jurkat (Human T-lymphoblastic cells) cell lysate Lane 4: HepG2 cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 30 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA721226, 1/1,000 in primary antibody, overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 186.0 kDa Observed band size: 250~300 kDa
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