C2C12 (cat. no. CRL-1772) was purchased from American Type Culture Collection. pCW-Cas9 and pLX-sgRNA were gifts from Eric Lander & David Sabatini (Addgene plasmid #50661 and #50662). Anti-LSD1 antibody (Cell Signaling Technology; cat. no. 2139), anti-beta-tubulin antibody (Millipore; cat. no. 05-661), anti-lamin B antibody (Santa Cruz; cat. no, SC-6217) and anti-RUNX2 antibody (MBL, cat. no. D130-3) were used for western blot. Anti-LSD1 antibody (Cell Signaling Technology; cat. no. 2184), anti-H3K4me1 (Cell Signaling Technology; cat. no. 5326), anti-H3K4me2 (Cell Signaling Technology; cat. no. 9725) and anti-H3K27ac (Cell Signaling Technology; cat. no. 8173) were used for ChIP analysis.
Anti lsd1 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-LSD1 antibody is a laboratory tool designed to detect the presence and localization of the LSD1 (Lysine-specific demethylase 1) protein in biological samples. LSD1 is an enzyme involved in the epigenetic regulation of gene expression. The antibody can be used in various analytical techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and distribution of LSD1 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti lamin b antibody, by Santa Cruz Biotechnology (1 mentions)
Anti h3k4me2, by Cell Signaling Technology (1 mentions)
Anti h3k4me1, by Cell Signaling Technology (1 mentions)
Anti h3k27ac, by Cell Signaling Technology (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Phosphatase inhibitor, by Merck Group (1 mentions)
M per, by Thermo Fisher Scientific (1 mentions)
Ecl prime western blotting system, by GE Healthcare (1 mentions)
Las 4000, by Cytiva (1 mentions)
Anti rap1 antibody, by Santa Cruz Biotechnology (1 mentions)
Catch and release co immunoprecipitation kit, by Merck Group (1 mentions)
Anti trf2 antibody, by Novus Biologicals (1 mentions)
4 protocols using anti lsd1 antibody
1
Characterizing C2C12 Myoblast Epigenetics
2
Western Blot Analysis of LSD1
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The nuclear extracts (4 µg protein equiv.) were denatured in 6× sample buffer (0.4 M Tris-HCl, pH 6.8, 12% SDS, 45% glycerol, 0.024% bromophenol blue and 10% 2-mercaptoethanol) by boiling at 95°C for 5 min, resolved by SDS-PAGE and transferred to a PVDF membrane. The membrane was incubated with a polyclonal anti-LSD1 antibody (1:1000, #2139, Cell Signaling Technology, Danvers, MA). Luminescence was produced by incubation with the SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific, Rockford, IL) and detected by using a LAS-1000 imaging system (GE Healthcare Japan, Tokyo, Japan). Collected images were analyzed by the ImageJ software (U.S. National Institutes of Health, Bethesda, MD).
3
Western Blot Analysis of LSD1 Protein
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The antibodies used for western blot analysis were anti-LSD1 antibody (1:1000, Cell Signaling Technology, Beverly, MA, USA) and mouse anti-GAPDH monoclonal antibody (1:3000, Wako, Osaka, Japan). Cells were lysed in M-PER (Thermo Scientific, Rockford, IL, USA) supplemented with protease inhibitors (Roche) and phosphatase inhibitors (Sigma-Aldrich, St. Louis, MO). Proteins were separated on a 4–20% polyacrylamide gradient-SDS gel, transferred on polyvinylidene difluoride membranes, and blocked in TBST (Tris-buffered saline and Tween20; 25 mM Tris, pH 7.4, 136 mM NaCl, 5 mM KCl, and 0.1% Tween) containing 5% milk. The antibodies were used at each dilution, as described above, in 5% milk/TBST. Blots were incubated with primary antibodies for 12 h at room temperature. Blots were washed (three times) with 5% milk/TBST and were incubated with the appropriate horseradish peroxidase-conjugated antibodies. Bound antibodies were detected with the ECL Prime Western Blotting System (RPN2232; GE Healthcare, Little Chalfont, Buckinghamshire, UK), and luminescent images were analyzed using a lumino imager (LAS-4000 mini; Fuji Film Inc. Tokyo, Japan).
4
TRF2, LSD1, and RAP1 Interactions
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HT-1080 cells were collected and washed in cold 1X PBS and nuclear extract was isolated using nuclear extract kit (Cell Extract from Sigma) as per manufacturer protocol. For immunoprecipitation experiments 500 µg of nuclear extract was incubated for 4 hours at 4 °C with 5 µg of anti-TRF2 antibody (Novus Biological) immunoprecipitation was performed using Catch and Release co-immunoprecipitation kit (Millipore) as per manufacturer’s protocol using anti-LSD1 antibody (Cell Signalling Technology) and anti-RAP1 antibody (Santacruz biotechnology). For reverse CoIP, 5 µg of LSD1 was used for immunoprecipitation.
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