Estrogen Receptor Alpha (N-Terminal) CUTANA™ CUT&RUN Antibody
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Powered by BiozThis antibody meets EpiCypher’s “CUTANA Compatible” criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. Estrogen Receptor Alpha N-terminal (ER alpha N-term) antibody shows CUT&RUN peaks in response to estradiol stimulation (Figure 1) that overlap with known estrogen response element (ERE) binding motifs (Figure 2). Overlap is further observed with peaks from an antibody to a different ER alpha epitope (C-term) and NCOA3 (SRC3), which co-activates ER-mediated transcription [1] (Figure 2).
Trademarks and copyrights
Background References:
[1] Wagner et al. BMC Cancer (2013). PMID: 24304549
Serum-starved MCF7 cells were treated with estradiol (E2) or vehicle control for 45 minutes. CUT&RUN was performed on 500k cells with 0.5 µg of either ER Alpha (N-Term), ER Alpha (C-Term; EpiCypher 13-2012), NOCA3/SRC3 (EpiCypher 13-2013), H3K4me3 positive control (EpiCypher 13-0041), or IgG negative control (EpiCypher 13-0042) antibodies using the CUTANA™ ChIC/CUT&RUN Kit v2.0 (EpiCypher 14-1048). Library preparation was performed with 5 ng of DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Both kit protocols were adapted for high throughput Tecan liquid handling. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2×50 bp). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.
Figure 1: ER alpha N-term peaks in CUT&RUN
Figure 2: ER alpha N-term peak analysis in CUT&RUN
Figure 3: Immunoprecipitation data
EpiCypher ER alpha N-term antibody (3 µg) was used to immunoprecipitate whole cell lysates isolated from MCF7 cells (1.0 mg per IP). A negative control IgG antibody and positive control antibody to different ER alpha epitopes (EpiCypher 13-2012 and Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto a 4-8% SDS-PAGE gel (25% of IP loaded) and probed via western blot with EpiCypher 13-2012 ER alpha C-term antibody (0.1 µg/mL).
Figure 4: Immunohistochemistry data
- Type: Polyclonal
- Host: Rabbit
- Applications: CUT&RUN, IP, IHC
- Reactivity: Human, Mouse (predicted), Rat (predicted)
- Format: Antigen affinity-purified
- Target Size: 66 kDa
Immunogen
Between amino acids 1 and 50
- CUT&RUN: 0.5 µg per reaction
- Immunoprecipitation (IP): 3- 5 µg/mg lysate
- Immunohistochemistry (IHC): 1:1,000 – 1:5,000. Epitope retrieval with citrate buffer pH 9.0 is recommended for FFPE tissue sections
- UniProt ID: P03372
- Gene Name: ESR1
- Protein Name: Estrogen receptor
- Alternate Names: ER, ER-alpha, Estradiol receptor, Nuclear receptor subfamily 3 group A member 1, ESR, NR3A1
