Anti mll1 antibody

Manufactured by Active Motif

Sourced in United States

The Anti-MLL1 antibody is a laboratory research tool that specifically recognizes and binds to the MLL1 (Mixed-Lineage Leukemia 1) protein. MLL1 is a histone methyltransferase involved in the regulation of gene expression. This antibody can be used to detect and study the MLL1 protein in various cellular and molecular biology applications.

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Lab products found in correlation

Anti tet1, by Thermo Fisher Scientific (2 mentions) Anti ezh2, by Cell Signaling Technology (2 mentions) Anti h3k4me3, by Cell Signaling Technology (1 mentions) Anti h3k27me3, by Cell Signaling Technology (1 mentions) Enhanced chemiluminescence western blotting detection kit, by Cytiva (1 mentions) Anti il 6, by Thermo Fisher Scientific (1 mentions) Anti myd88, by Cell Signaling Technology (1 mentions) Anti dnmt1, by Abcam (1 mentions) Anti il 1β, by Thermo Fisher Scientific (1 mentions) Ab171870, by Abcam (1 mentions) Bioruptor pico, by Diagenode (1 mentions) Anti h3k4me3 antibody, by Active Motif (1 mentions) Anti p16ink4a, by Cell Signaling Technology (1 mentions) Anti dnmt1 antibody, by Abcam (1 mentions) Anti ahr, by Thermo Fisher Scientific (1 mentions)

3 protocols using anti mll1 antibody

Protein Expression Analysis Protocol

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Crude protein in cell lysates was electrophoresed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride membrane that was incubated with primary antibodies. Anti-IL-1β, anti-IL-6, and anti-TET1 antibodies were purchased from Thermo Fisher Scientific; anti-TLR5, anti-phospho-NFκB (p65) and anti-DNMT1 antibodies were from Abcam (Cambridge, UK); anti-MyD88, anti-EZH2, anti-H3K27Me3, and anti-H3K4Me3 antibodies were from Cell Signaling Technology (Danvers, MA, USA); anti-NOX4 and anti-DNMT3B antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and anti-MLL1 antibody was from Active Motif (Carlsbad, CA, USA). Then, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Pierce, Rockland, IL, USA), and protein bands were detected using an enhanced chemiluminescence western blotting detection kit (Amersham, Little Chalfont, Buckinghamshire, UK).

Ryu Y.S., Kang K.A., Piao M.J., Ahn M.J., Yi J.M., Hyun Y.M., Kim S.H., Ko M.K., Park C.O, & Hyun J.W. (2018). Particulate matter induces inflammatory cytokine production via activation of NFκB by TLR5-NOX4-ROS signaling in human skin keratinocyte and mouse skin. Redox Biology, 21, 101080.

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ChIP-qPCR Analysis of H3K4me3 and MLL1

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As previously described (48 (link)), cells were fixed in 1% paraformaldehyde and glycine. This was followed by lysis and sonication using a Bioruptor Pico (Diagenode) to generate 300–500 bp fragments. Samples were then incubated overnight with anti-H3K4me3 antibody (39159, Active Motif), anti-MLL1 antibody (61296, Active Motif), or isotype control (rabbit polyclonal IgG; ab171870, Abcam), followed by the addition of protein A or protein G Sepharose beads (Thermo Fisher Scientific). Beads were washed; then, DNA was eluted and purified using phenol/chloroform/isoamyl alcohol extraction. The DNA was then precipitated using ethanol. H3K4me3 deposition was measured by qPCR using 2× SYBR PCR mix (Invitrogen), and primers (forward, 5′-TCTCTGACACCTTTGCCTGA-3′; reverse, 5′-CCAGTTCTCTTCCCTTGCAC-3′) were designed to flank the Ifnk promoter using NCBI Primer-BLAST.

Wolf S.J., Audu C.O., Joshi A., denDekker A., Melvin W.J., Davis F.M., Xing X., Wasikowski R., Tsoi L.C., Kunkel S.L., Gudjonsson J.E., O’Riordan M.X., Kahlenberg J.M, & Gallagher K.A. (2022). IFN-κ is critical for normal wound repair and is decreased in diabetic wounds. JCI Insight, 7(9), e152765.

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Western Blot Analysis of Epigenetic Regulators

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Cell lysates (50 μg protein) were electrophoresed, transferred, and immunoblotted with specific antibodies. Anti-AhR, anti-TET1, and anti-TBP antibodies were purchased from Thermo Fisher Scientific, Inc.; the anti-DNMT1 antibody was from Abcam (Burlingame, CA, USA); the anti-DNMT3B antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA); the anti-p16^INK4A and anti-EZH2 antibodies were from Cell Signaling Technology, Inc. (Danvers, MA, USA), and the anti-MLL1 antibody was purchased from Active Motif (Carlsbad, CA, USA). The membranes with bound primary antibodies (1:1000) were reacted with horseradish peroxidase (HRP)-conjugated secondary antibodies (Pierce, Rockland, IL, USA), and the protein bands were assessed by a western blotting detection kit (Amersham, Little Chalfont, Buckinghamshire, UK).

Ryu Y.S., Kang K.A., Piao M.J., Ahn M.J., Yi J.M., Bossis G., Hyun Y.M., Park C.O, & Hyun J.W. (2019). Particulate matter-induced senescence of skin keratinocytes involves oxidative stress-dependent epigenetic modifications. Experimental & Molecular Medicine, 51(9), 108.

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