The following antibodies were used in this study: anti-H3K4me1 (generated in-house), anti-H3K4me2 (generated in-house), anti-H3K4me3 (generated in-house), anti-H3K27ac (Cell Signaling, 8173), anti-H3 (generated in-house), anti-SMC1A (Bethyl Laboratories, A300-055A), anti-SET1A (generated in-house), anti-MLL1 (Cell Signaling, 14689), anti-MLL2 (generated in-house), anti-MLL3 (generated in-house), anti-MLL4 (generated in-house), anti-RBBP5 (Bethyl Laboratories, A300-109A), anti-HSP90 (Santa Cruz Biotechnology, 7947), and anti-tubulin (Developmental Studies Hybridoma Bank, E7).
Anti rbbp5
Manufactured by Fortis Life Sciences
Anti-RBBP5 is a laboratory reagent used for detecting and analyzing the RBBP5 protein in biological samples. RBBP5 is a core component of the Set1/MLL histone methyltransferase complex, which is involved in the regulation of gene expression. The anti-RBBP5 reagent can be used in various experimental techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of the RBBP5 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti ash2l, by Fortis Life Sciences (6 mentions)
Anti h3k4me1, by Cell Signaling Technology (3 mentions)
Anti mll2, by Fortis Life Sciences (3 mentions)
Anti wdr5, by Fortis Life Sciences (3 mentions)
Anti h3k4me3, by Abcam (3 mentions)
Anti h3k4me3, by Cell Signaling Technology (2 mentions)
Anti h3k27ac, by Cell Signaling Technology (2 mentions)
Anti tubulin, by Developmental Studies Hybridoma Bank (2 mentions)
Anti flag m2, by Merck Group (2 mentions)
Anti flag, by Merck Group (2 mentions)
Anti mll1c, by Fortis Life Sciences (2 mentions)
Cycletest plus dna reagent kit, by BD (2 mentions)
Alexa fluor goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Anti tubulin, by Merck Group (2 mentions)
Alexa fluor 680 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions)
Anti apaf1, by R&D Systems (2 mentions)
Anti pig3, by Abcam (2 mentions)
Anti p21, by BD (2 mentions)
Anti gapdh, by Merck Group (2 mentions)
Anti hsp90, by Santa Cruz Biotechnology (1 mentions)
13 protocols using anti rbbp5
1
Comprehensive Histone Modification Profiling
2
Epigenetic Markers Immunostaining Protocol
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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]).
3
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).
4
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).
5
Protein Extraction and Immunostaining in C2C12 Cells
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The total proteins were extracted using RIPA lysis buffer. The following dilutions of antibodies were used for each antibody: anti-MyoG (1:2000, sc-576, Santa Cruz), anti-MyHC (1:2000, M4276, Sigma), anti-hnRNPL (1:5000, sc-28726, Santa Cruz), anti-hnRNPK (1:5000, 4675, Cell Signaling), anti-MED1 (1:5000, A300–793A, Bethyl Laboratories), anti-RAD21 (1:5000, A300–080A, Bethyl Laboratories), anti-RBBP5 (1:5000, A300–109A, Bethyl Laboratories), anti-YY1 (1:2000, sc-1703, Santa Cruz), anti-MyoD (1:2000, sc-760, Santa Cruz), anti-α-Tubulin (1:5000, sc-23948, Santa Cruz), and anti-H3K36me3 (1:5000, ab9050, Abcam). For Immunofluorescence staining of cultured C2C12 cells, the following dilutions were used: anti-MyHC (1:350, M4276, Sigma). All fluorescent images were captured with a Nikon fluorescence microscope.
6
Histone Modification Antibody Panel
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The following antibodies are used in this study: anti-H3K4me1 (Cell Signaling Technology [CST], 5326), anti-H3K4me2 (generated in house), anti-H3K4me3 (CST, 9751), anti-H3K27ac (CST, 8173), H3K27me3 (CST, 9733), anti-MLL3 NT (generated in house), anti-MLL3 MT (generated in house), anti-MLL4 NT (generated in house), anti-MLL4 CT (generated in house), anti-MLL2 (generated in house), anti-SET1A (generated in house), anti-Menin (CST, 6891), anti-MLL1C (CST, 14197), anti-RBBP5 (Bethyl Laboratories, A300-109A), anti-NCOA6 (Bethyl Laboratories, A300-410A), anti-UTX (CST, 33510), anti-PTIP (Bethyl Laboratories, A300-370A), ASH2L (CST, 5019), H3 Ser10-p (CST 53348), CDT1 (CST, 8064), Cyclin B1 (CST, 12231), Geminin (CST, 52508), Cyclin E1 (CST, 20808), Cyclin A2 (CST, 91500), p-cdc2 (CST, 4539), PARP (CST, 9542) Caspase3 (CST, 9662), anti-GART (Santa Cruz Biotechnology, sc-166447), anti-PAICS (Proteintech, 12967-1-AP), Hsp90 (Santa Cruz Biotechnology, sc-13119), and anti-β-tubulin (Developmental Studies Hybridoma Bank, E7). Western blot analysis was performed as previously described (53 (link)).
7
Western Blot Analysis of MLL Complexes
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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.
8
Antibody Validation for Protein Interactions
9
Immortalized Cell Lines for Chromatin Studies
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HEK293T/17 cells were obtained from the American Type Culture Collection (ATCC). The HEK293T 5XUAS-Luciferase cell line has been previously described (Wysocka et al., 2005 (link)). V6.5 mouse embryonic stem cells were grown on 0.2% gelatinized plates in Knockout DMEM supplemented with 15% FBS, 1% Glutamax (35050; Invitrogen, Carlsbad, CA), 1% nonessential amino acids, 1% Pen/Strep, 0.2% β-mercaptoethanol, LIF (ESGRO1107; 1:10000; Millipore, Billerica, MA), and 2 µg/ml doxycycline (D9891; Sigma, St. Louis, MO). To maintain consistent levels of doxycycline, media was changed every 2 days. Antibodies used were: anti-FLAG-M2 (F1804; Sigma, St. Louis, MO), anti-β actin (ab8227; Abcam, Cambridge, MA), anti-RbBP5 (A300-109A; Bethyl, Montgomery, TX), anti-H3K4me3 (ab8580; Abcam, Cambridge, MA), anti-β tubulin (ab6046; Abcam, Cambridge, MA), anti-H3 (ab1791; Abcam, Cambridge, MA), anti-WDR5 (07-706; Millipore, Billerica, MA), and anti-HA (MMS-101P; Covance, Princeton, NJ). All immunoprecipitations were conducted using anti-FLAG-M2 (A2220; Sigma, St. Louis, MO) or mouse IgG (A0919; Sigma, St. Louis, MO) agarose beads. Unless noted, all expression vectors were cloned into pcDNA3 or pcDNA3.1 + backbones.
10
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.
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