ChIP-seq was performed by Active Motif (Carlsbad, CA). Briefly, 1×106 MCF7 cells were seeded in 15cm dishes in phenol red-free medium supplemented with 5% CSS for 72 hrs. Cells were pre-treated with vehicle, 10 µM Enza, or 30uM MJC13 for 3 hrs. E2 was then added for 1 hr in continued presence of vehicle, Enza, or MJC13. For AR ChIP-seq, an additional sample was treated with DHT for 4 hrs. The cells were washed with PBS then fixed as per manufacturer’s instructions (Active Motif). AR antibody H-280 (Santa Cruz) or ER antibody HC-20 (Santa Cruz) were utilized. Peak calls were made by MACS2 (23 (link)) with default parameters using the sequence alignments obtained from Active Motif. Motif discovery was performed on 100 base pairs surrounding the peak summit using BioProspector (24 (link)). Patser (25 (link)) was used to determine significant matches to AREs and EREs.
Ar antibody h 280
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The AR antibody H-280 is a rabbit polyclonal antibody that recognizes the androgen receptor (AR) protein. It is designed for use in various research applications, such as Western blotting, immunohistochemistry, and immunoprecipitation, to detect and study the AR protein.
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Lab products found in correlation
2 protocols using ar antibody h 280
1
ChIP-seq Analysis of AR and ER
2
Integrating ChIP-seq and Cell Culture Techniques for Transcription Factor Analysis
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For AR ChIP-seq, MDA-MB-453 cells were grown in charcoal-stripped serum media for a total of 72 h before treatment. Twenty-four hours prior to treatment, cells were trypsinized and equal cell numbers were plated on control tissue culture dishes (attached) or poly-HEMA coated dishes (suspended). Cells were treated with DMSO (vehicle control), DHT (10 nM), or DHT + Enza (10 µM) for 4 h, followed by fixation in 1% formaldehyde. ChIP-seq was performed as previously reported [34 (link)]. Chromatin was sonicated using an EpiShear Probe Sonicator (Active Motif, Carlsbad, CA, USA) for 4 min (cycles of 30 s with 30 s of rest in between) at 40% power. AR antibody H-280 (Santa Cruz Biotechnology) was utilized for immunoprecipitation and libraries were sequenced on an HiSeq 2500 (Illumina, San Diego, CA, USA).
RUNX1 ChIP in an MDA-MB-231 cell line was performed as previously described [22 (link)] using anti-RUNX1 (Abcam, Cambridge, UK; #23980) and anti-IgG (Abcam; #46540, negative control). Primer sequences are available insupplementary Table S4b .
RUNX1 ChIP in an MDA-MB-231 cell line was performed as previously described [22 (link)] using anti-RUNX1 (Abcam, Cambridge, UK; #23980) and anti-IgG (Abcam; #46540, negative control). Primer sequences are available in
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