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Kmt2a mll1 complex

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The KMT2A (MLL1) complex is a protein complex involved in the regulation of gene expression. It possesses histone methyltransferase activity, which means it can add methyl groups to histone proteins, thereby modifying the chromatin structure and influencing gene transcription.

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Lab products found in correlation

Bl21 star de3 cells, by Thermo Fisher Scientific (2 mentions) Recombinant human tgm2, by R&D Systems (2 mentions) Fibrinogen from human plasma, by Merck Group (2 mentions)

2 protocols using kmt2a mll1 complex

1

Purification of Recombinant Histones

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Human histones H3.2 and H4 were cloned into pRUTH5 to yield an N-terminal His6-TEV tag. Histones were then expressed in BL21 Star(DE3) cells (Thermo Fisher, cat#C6010–03) into inclusion bodies for 3  hr at 37°C. Inclusion bodies were re-solubilized in 6  M guanidine-HCl, 1  M NaCl and 50  mM Tris-HCl (pH  8) prior to purification on a Ni-NTA affinity column. To generate recombinant H3Q5A, mutations were introduced into the pRUTH5-H3.2 construct using standard SDM. Additional histones were purchased from Active Motif (H3.1, cat#31294 and EPL generated H3K4me3, cat#31278). For some experiments, recombinant human TGM2 was purchased from R&D Systems (cat#4376-TG-050), and guinea pig Tgm2 was purchased from Zedira (cat#T006); KMT2A (MLL1) complex from Active Motif (cat#31423); and Fibrinogen from human plasma from Sigma (cat#F4883).
Farrelly L.A., Thompson R.E., Zhao S., Lepack A.E., Lyu Y., Bhanu N.V., Zhang B., Loh Y.H., Ramakrishnan A., Vadodaria K.C., Heard K.J., Erikson G., Nakadai T., Bastle R.M., Lukasak B.J., Zebroski H I.I.I., Alenina N., Bader M., Berton O., Roeder R.G., Molina H., Gage F.H., Shen L., Garcia B.A., Li H., Muir T.W, & Maze I. (2019). Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3. Nature, 567(7749), 535-539.
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2

Purification of Recombinant Histones

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human histones H3.2 and H4 were cloned into pRUTH5 to yield an N-terminal His6-TEV tag. Histones were then expressed in BL21 Star(DE3) cells (Thermo Fisher, cat#C6010–03) into inclusion bodies for 3  hr at 37°C. Inclusion bodies were re-solubilized in 6  M guanidine-HCl, 1  M NaCl and 50  mM Tris-HCl (pH  8) prior to purification on a Ni-NTA affinity column. To generate recombinant H3Q5A, mutations were introduced into the pRUTH5-H3.2 construct using standard SDM. Additional histones were purchased from Active Motif (H3.1, cat#31294 and EPL generated H3K4me3, cat#31278). For some experiments, recombinant human TGM2 was purchased from R&D Systems (cat#4376-TG-050), and guinea pig Tgm2 was purchased from Zedira (cat#T006); KMT2A (MLL1) complex from Active Motif (cat#31423); and Fibrinogen from human plasma from Sigma (cat#F4883).
Farrelly L.A., Thompson R.E., Zhao S., Lepack A.E., Lyu Y., Bhanu N.V., Zhang B., Loh Y.H., Ramakrishnan A., Vadodaria K.C., Heard K.J., Erikson G., Nakadai T., Bastle R.M., Lukasak B.J., Zebroski H I.I.I., Alenina N., Bader M., Berton O., Roeder R.G., Molina H., Gage F.H., Shen L., Garcia B.A., Li H., Muir T.W, & Maze I. (2019). Histone serotonylation is a permissive modification that enhances TFIID binding to H3K4me3. Nature, 567(7749), 535-539.
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