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Anti ash2l

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Anti-ASH2L is a laboratory reagent that targets the ASH2L protein. ASH2L is a component of the Set1/MLL histone methyltransferase complex, which is involved in the regulation of gene expression. Anti-ASH2L can be used to study the function and localization of the ASH2L protein in biological systems.

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Lab products found in correlation

Anti rbbp5, by Fortis Life Sciences (6 mentions) Anti wdr5, by Fortis Life Sciences (3 mentions) Anti h4k20me1, by Active Motif (2 mentions) Anti flag, by Merck Group (2 mentions) Anti apaf1, by R&D Systems (2 mentions) Anti pig3, by Abcam (2 mentions) Alexa fluor goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Alexa fluor 680 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Anti tubulin, by Merck Group (2 mentions) Anti h3k4me3, by Abcam (2 mentions) Cycletest plus dna reagent kit, by BD (2 mentions) Anti p21, by BD (2 mentions) Anti ring1b, by Cell Signaling Technology (1 mentions) Anti flag m2, by Merck Group (1 mentions) Anti ezh2, by BD (1 mentions) Anti trimethyl histone h3 lys4, by Merck Group (1 mentions) Anti lana ln53, by Merck Group (1 mentions) Goat anti rat igg h, by Thermo Fisher Scientific (1 mentions) Anti mll1c, by Fortis Life Sciences (1 mentions) Anti t7 tag antibody, by Abcam (1 mentions)

8 protocols using anti ash2l

1

Epigenetic Markers Immunostaining Protocol

Cited 1 time
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Immuno-FISH was performed as described [28 (link),67 (link)]. Antibodies used in this study are the following: anti-H3K27me3 (#0323, MABI [1:1000 dilution] and #9733, Cell Signaling [1:1000]), anti-uH2A (#8240, Cell Signaling [1:1000]), anti-H4K20me1 (#39727, Active motif [1:5000]), anti-RBBP5 (A300-109A, Bethyl Laboratories [1:400]), anti-ASH2L (A300-107A, Bethyl Laboratories [1:400]), anti-EZH2 (#612666, BD [1:200]), anti-RING1B (#5694, Cell Signaling [1:50]) and anti-FLAG-M2 (F1804, SIGMA [1:4000]).
Yue M., Ogawa A., Yamada N., Charles Richard J.L., Barski A, & Ogawa Y. (2017). Xist RNA repeat E is essential for ASH2L recruitment to the inactive X and regulates histone modifications and escape gene expression. PLoS Genetics, 13(7), e1006890.
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2

Chromatin Immunoprecipitation (ChIP) Assay

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Antibodies used included anti-Flag (Sigma, F3165-5MG, clone M2), anti-MLL1C (Bethyl, A300-374A), anti-RbBP5 (Bethyl, A300-109A), anti-ASH2L (Bethyl, A300-112A), anti-WDR5 (Bethyl, A302-430A), anti-GFP (Clontech, 632377, clone JL-8), anti-trimethyl-Histone H3 (Lys4) (EMD Millipore, 07-473), anti-LANA LN53 (Millipore, MABE1109), goat anti-rat IgG (H + L) Alexa Fluor 488 (Invitrogen A11006), goat anti-rabbit IgG (H + L) Alexa Fluor 647 (Invitrogen A21245), human anti -LANA sera adsorbed against uninfected cell extract for western blot or affinity purified against carboxy-terminal LANA for ChIP, Anti-T7 tag® antibody (Abcam, ab9138), and anti-alpha-tubulin (Sigma T9026, cline DM1A).
Tan M., Li S., Juillard F., Chitas R., Custódio T.F., Xue H., Szymula A., Sun Q., Liu B., Álvarez Á.L., Chen S., Huang J., Simas J.P., McVey C.E, & Kaye K.M. (2021). MLL1 is regulated by KSHV LANA and is important for virus latency. Nucleic Acids Research, 49(22), 12895-12911.
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3

Comprehensive Western Blotting and Cell Cycle Analysis

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Western blotting was performed as described previously 8 (link), 61 (link), with the exception that Alexa Fluor secondary antibodies were used. All blots were developed using the Odyssey fluorescence image scanner and the band intensities were quantified using LI–COR software. Cell cycle analysis was performed using PI (propidium iodide) staining as described previously 8 (link), using the CycleTEST PLUS DNA reagent kit (BD Biosciences, San Jose, CA, USA; # 340242). The following antibodies were used in the study: anti–P53 (SCBT; sc–6243), anti–P21 (BD; #556431), anti–APAF1 (R & D; #MAB828), anti–PIG3 (abcam; #ab64798), anti–PUMA (CST; #4976); anti–Ash2L (Bethyl Labs; #A300-108A), anti–H3K4me3 (abcam; #ab1012), anti–Wdr5 (abcam; #ab56919); anti–RbBP5 (Bethyl Labs; #A300-109A), anti–RNAP II (SCBT; sc–899); anti–Serine5–CTD (abcam; #ab5131–50); anti–TAFII (SCBT; sc–735); anti–TFIIB (SCBT; sc–225); anti–TFIIF (SCBT; sc–235); anti–Tubulin (Sigma; #T5168), Alexa Fluor 680 goat anti–mouse IgG (Molecular Probes; #A21057) and Alexa Fluor goat anti–rabbit IgG (Molecular Probes; #A21076).
Mungamuri S.K., Wang S., Manfredi J.J., Gu W, & Aaronson S.A. (2014). Ash2L enables P53–dependent apoptosis by favoring stable transcription pre–initiation complex formation on its pro-apoptotic target promoters. Oncogene, 34(19), 2461-2470.
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4

Antibody Validation for Protein Interactions

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Antibodies are as follows: anti-HA (Abcam, ab9110), anti-AKAP95 (Santa Cruz Biotechnology, sc-10766), anti-His (Santa Cruz Biotechnology, sc-803), anti-GAPDH (Chemicon, MAB374), anti-FLAG (Sigma, A8592, for blotting), anti-FLAG [M2 beads] (Sigma, A2220), and anti-H3K4me3 (Millipore, 07–473), anti-RBBP5 (Bethyl Laboratories, A300–109A), anti-ASH2L (Bethyl Laboratories, A300–107A), anti-WDR5 (Bethyl Laboratories, A302–430A).
Shah K.K., Whitaker R.H., Busby T., Hu J., Shi B., Wang Z., Zang C., Placzek W.J, & Jiang H. (2019). Specific inhibition of DPY30 activity by ASH2L-derived peptides suppresses blood cancer cell growth. Experimental cell research, 382(2), 111485.
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5

Chromatin Immunoprecipitation (ChIP) Assay

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Chromatin immunoprecipitation (ChIP) assays were performed as described previously (18 (link)). Rabbit polyclonal antibodies for ChIP were either custom generated for EBNA1 (Pocono Rabbit Farm) or were purchased for rabbit and mouse anti-IgG (Santa Cruz Biotechnology), anti-HCF1 (catalog no. A301-400A; Bethyl), anti-RbBp5 (catalog no. A300-109A; Bethyl), anti-Ash2L (catalog no. A300-489A; Bethyl), anti-OCT2 (catalog no RB9297; Neo Markers) and pan-H3 (catalog no. 07-690), H3K4me3 (catalog no. 07-473), H3K9acetyl (catalog no. 07-352) were purchased from Millipore. Rabbit serum anti-H3K9me3 (catalog no. 39161), anti-H3K4me1 (catalog no. 61633), anti-H3K27me3 (catalog no. 39155)- and anti-H3K27acetyl (catalog no. 39685), and anti-H3K4me2 (catalog no. 39141) were from Active Motif.
Dheekollu J., Wiedmer A., Sentana-Lledo D., Cassel J., Messick T, & Lieberman P.M. (2016). HCF1 and OCT2 Cooperate with EBNA1 To Enhance OriP-Dependent Transcription and Episome Maintenance of Latent Epstein-Barr Virus. Journal of Virology, 90(11), 5353-5367.
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6

Comprehensive Western Blotting and Cell Cycle Analysis

Cited 1 time
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Western blotting was performed as described previously 8 (link), 61 (link), with the exception that Alexa Fluor secondary antibodies were used. All blots were developed using the Odyssey fluorescence image scanner and the band intensities were quantified using LI–COR software. Cell cycle analysis was performed using PI (propidium iodide) staining as described previously 8 (link), using the CycleTEST PLUS DNA reagent kit (BD Biosciences, San Jose, CA, USA; # 340242). The following antibodies were used in the study: anti–P53 (SCBT; sc–6243), anti–P21 (BD; #556431), anti–APAF1 (R & D; #MAB828), anti–PIG3 (abcam; #ab64798), anti–PUMA (CST; #4976); anti–Ash2L (Bethyl Labs; #A300-108A), anti–H3K4me3 (abcam; #ab1012), anti–Wdr5 (abcam; #ab56919); anti–RbBP5 (Bethyl Labs; #A300-109A), anti–RNAP II (SCBT; sc–899); anti–Serine5–CTD (abcam; #ab5131–50); anti–TAFII (SCBT; sc–735); anti–TFIIB (SCBT; sc–225); anti–TFIIF (SCBT; sc–235); anti–Tubulin (Sigma; #T5168), Alexa Fluor 680 goat anti–mouse IgG (Molecular Probes; #A21057) and Alexa Fluor goat anti–rabbit IgG (Molecular Probes; #A21076).
Mungamuri S.K., Wang S., Manfredi J.J., Gu W, & Aaronson S.A. (2014). Ash2L enables P53–dependent apoptosis by favoring stable transcription pre–initiation complex formation on its pro-apoptotic target promoters. Oncogene, 34(19), 2461-2470.
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7

Protein Interaction and Transcriptional Regulation Assays

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All plasmids about BAP18 were introduced in our previous work (39 (link)). ERα expression plasmid was cloned into pSG5 plasmid and 3 estrogen response elements were cloned made into pGL-ERE-adML reporter plasmid to perform luciferase reporter assay.
The antibodies were used in our study as follow: anti-BAP18 (Bethyl #A304-207A-1), anti-ERα (Cell signaling #D8H8), anti-MYC (Thermo Fisher #9E10), anti-CyclinD1 (Cell signaling #DCS6), anti-GAPDH (Kangchen #KC5G4), anti-WDR5 (Bethyl #A302-429A-2), anti-ASH2L (Bethyl #A300-107A-2), anti-DPY30 (Abcam #ab187690), anti-H3K4me3/H3ac/H3K9ac/H4ac/H4K16ac (Millipore), anti-Rabbit/Mouse (ABclonal), anti-IgG (Proteintech#10238-1-AP), anti-GFP (Sigma #G1544) and anti-FLAG (Proteintech#20543-1-AP).
Sun G., Wang C., Wang S., Sun H., Zeng K., Zou R., Lin L., Liu W., Sun N., Song H., Liu W., Zhou T., Jin F., Shan Z, & Zhao Y. (2020). An H3K4me3 reader, BAP18 as an adaptor of COMPASS-like core subunits co-activates ERα action and associates with the sensitivity of antiestrogen in breast cancer. Nucleic Acids Research, 48(19), 10768-10784.
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8

Immuno-FISH Protocol for Histone Modifications

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Immuno-FISH was performed as described (21 (link),35 (link)). Antibodies used in this study are: anti-H3K27me3 (MABI #0323, 1:1000 dilution and Cell Signaling #9733, 1:1000), anti-H2AK119Ub (Cell Signaling #8240, 1:1000), anti-H4K20me1 (Active motif #39727, 1:5000), anti-ASH2L (Bethyl Laboratories A300-107A, 1:400).
Yue M, & Ogawa Y. (2017). CRISPR/Cas9-mediated modulation of splicing efficiency reveals short splicing isoform of Xist RNA is sufficient to induce X-chromosome inactivation. Nucleic Acids Research, 46(5), e26.
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