Kmt1a

Manufactured by Cell Signaling Technology

KMT1A is a histone-lysine N-methyltransferase that catalyzes the mono-, di-, and tri-methylation of histone H3 at lysine 9 (H3K9). It plays a crucial role in the epigenetic regulation of gene expression and chromatin organization.

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Lab products found in correlation

β actin peroxidase, by Merck Group (2 mentions) Myogenin, by BD (1 mentions) Myosin heavy chain, by Developmental Studies Hybridoma Bank (1 mentions) Plko 1 trc vector, by Addgene (1 mentions) Ha peroxidase, by Merck Group (1 mentions) Trimethyl histone h3 lys 9, by Merck Group (1 mentions) Trimethyl histone h3 lys27, by Merck Group (1 mentions) Total p38, by Cell Signaling Technology (1 mentions) Phospho p38, by Cell Signaling Technology (1 mentions) P21cip1, by Santa Cruz Biotechnology (1 mentions) Normal rabbit igg, by Santa Cruz Biotechnology (1 mentions) Acetyl histone h3, by Merck Group (1 mentions) Flag m2, by Merck Group (1 mentions) Phospho h2ax ser139, by Cell Signaling Technology (1 mentions) Fluorchem hd2, by Bio-Techne (1 mentions) Flag m2 hrp, by Merck Group (1 mentions) Chemidoc touch, by Bio-Rad (1 mentions) Vectashield mounting medium, by Vector Laboratories (1 mentions)

2 protocols using kmt1a

Lentiviral Constructs for KMT1A and p38 Studies

Cited 1 time

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Lentiviral pLV vector expressing Flag-KMT1A [38 (link)] and LV-HA-MKK6EE and pLV-HA-MKK6DN were generated by subcloning inserts from pcDNA-HA-MKK6EE and pcDNA-HA-MKK6DN (provided by Dr. L. Puri) [39 (link)] into pLV vector. For expression of shRNA, KMT1A, p38α, or scramble shRNAs are cloned individually into lentiviral pLKO.1-TRC vector (Addgene) and sequence verified. The shRNA sequences for KMT1A and scramble were described previously [38 (link)]. The sequence for p38α shRNA was 5′-AGCCCAGCAACCTAGCTGTTT-3′. Vectors pGEX-4T-3-H3(N) [26 (link)] and pGEX-ATF2 (provided by Dr. J. Han) [40 (link)] express GST fusion N-terminal histone H3 and ATF2 proteins, respectively.
Antibodies used were phospho-p38 (Cell Signaling 9215), β-actin-peroxidase (Sigma A3854), Flag-M2 (Sigma F3165), myogenin (BD Pharmingen 556358), KMT1A (Cell Signaling 8729, and Millipore 07-550 and 05-615), MyoD (Santa Cruz sc-760 and BD Pharmingen 554130), p38α (Cell Signaling 9790), HA-peroxidase (Sigma H6533), acetyl-histone H3 (Millipore, 06-599), trimethyl-histone H3 (Lys-9) (Millipore 07-442), trimethyl-histone H3 (Lys27) (Millipore 07-449), GAPDH (Biodesign H86504M), Brg-1 (Santa Cruz sc-10768), p21^cip1 (Santa Cruz sc-397), myosin heavy chain (Developmental Studies Hybridoma Bank, MF-20), total p38 (Cell Signaling 9212), and normal rabbit IgG (Santa Cruz sc-2027).

Chatterjee B., Wolff D.W., Jothi M., Mal M, & Mal A.K. (2016). p38α MAPK disables KMT1A-mediated repression of myogenic differentiation program. Skeletal Muscle, 6, 28.

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Immunoblot and Immunofluorescence Protocols

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Preparation of cell extracts and immunoblot analyses were carried out as described previously [46 (link)]. Immunofluorescence was carried out as described previously [10 (link)], with VECTASHIELD mounting medium (Vector Labs) used to counterstain with DAPI. Antibodies used were β-Actin-peroxidase (Sigma A3854), Flag-M2-HRP (Sigma A8592), MyoD (Santa Cruz sc-304), MyoG (BD Pharmingen 556358), KMT1A (Cell Signaling 8729), total histone H2A (Cell Signaling L88A6), phospho-H2AX (Ser 139) (Cell Signaling 20E3), and MyHC (Developmental Studies Hybridoma Bank, MF-20). Images were generated using the FluorChem HD2 (Alpha Innotech) or ChemiDoc Touch (Bio-Rad) imaging systems, which produce high contrast images and can identify over-exposed protein bands.

Wolff D.W., Lee M.H., Jothi M., Mal M., Li F, & Mal A.K. (2018). Camptothecin exhibits topoisomerase1-independent KMT1A suppression and myogenic differentiation in alveolar rhabdomyosarcoma cells. Oncotarget, 9(40), 25796-25807.

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