Western Blot Analysis of MLL Complexes

Western blot analysis was performed as described previously (Zhao et al. 2013 (link)). Anti-TFIIA and anti-RbBP5 antibodies were purchased from Bethyl Laboratories. Anti-taspase1 was from Thermo Fisher Scientific. Anti-MLL2N and MLL2C antibodies and anti-histone H3, H3K4me1, H3K4me2, and H3K4me3 antibodies were homemade and validated in our previous studies (Hu et al. 2013 (link); Rickels et al. 2016 (link)). Anti-B-tubulin (E7) was purchased from the Developmental Studies Hybridoma Bank. Anti-Hsp90 antibodies were purchased from Santa Cruz Biotechnology. Menin (no. 6891), LEDGF (no. 2088), MLL1N (no. 14689), MLL1C (no. 14197), ASH2L (no. 5019), PARP (no. 9542), and p21 (no. 2947) antibodies were purchased from Cell Signaling Technology.

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Zhao Z., Wang L., Volk A.G., Birch N.W., Stoltz K.L., Bartom E.T., Marshall S.A., Rendleman E.J., Nestler C.M., Shilati J., Schiltz G.E., Crispino J.D, & Shilatifard A. (2019). Regulation of MLL/COMPASS stability through its proteolytic cleavage by taspase1 as a possible approach for clinical therapy of leukemia. Genes & Development, 33(1-2), 61-74.

Publication 2019

anti-RbBP5

Anti antibodies Antibodies H3k4me3 Histone h3 Hsp90 Hybridoma Ledgf Menin Tfiia Tubulin Western blot analysis

Corresponding Organization :

Other organizations : Northwestern University

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Variable analysis

independent variables

Western blot analysis

dependent variables

Expression levels of TFIIA, RbBP5, taspase1, MLL2N, MLL2C, histone H3, H3K4me1, H3K4me2, H3K4me3, B-tubulin, Hsp90, Menin, LEDGF, MLL1N, MLL1C, ASH2L, PARP, and p21

control variables

Antibodies used for Western blot analysis, including anti-TFIIA, anti-RbBP5, anti-taspase1, anti-MLL2N, anti-MLL2C, anti-histone H3, anti-H3K4me1, anti-H3K4me2, anti-H3K4me3, anti-B-tubulin (E7), anti-Hsp90, anti-Menin, anti-LEDGF, anti-MLL1N, anti-MLL1C, anti-ASH2L, anti-PARP, and anti-p21

positive controls

Not specified

negative controls

Not specified

Annotations

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