CUTANA™ Anti-GST Tag Antibody

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Figure 2: Gene browser tracks
meCUT&RUN was performed as described in Figure 4. A 30 kb window at the AJM1 gene is shown for anti-GST antibody and meCUT&RUN. Tracks are also shown with representative data for meCUT&RUN followed by EM-seq (meCUT&RUN-EM) and whole genome EM-seq (WGEM), using the New England Biolabs NEBNext^® Enzymatic Methyl-seq v2 Kit (NEB E8015). The meCUT&RUN kit produced the expected genomic distribution, showing enrichment of methylated DNA that approximates the methylated CpG pattern observed in WGEM. Images were generated using the Integrative Genomics Viewer (IGV, Broad Institute).

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Figure 3: Western blot data
Western analysis of GST Tag in 0.1, 0.3, or 1.0 µg of HEK293 cell lysate expressing a GST-tag fusion protein. Lysate was resolved via SDS-PAGE and detected with a 1:7,500 dilution of EpiCypher anti-GST Tag antibody.

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Figure 4: meCUT&RUN methods
meCUT&RUN was performed starting with 500k K562 cells with either 2.5 µL of 20X GST-MeCP2 (EpiCypher 15-2002) added as the primary binding reagent or 0.5 µg of a secondary antibody-only control (anti-GST antibody) to determine background cleavage. Library preparation was performed using 5 ng of meCUT&RUN-enriched DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 7.7 million reads (anti-GST) and 8.4 million reads (GST-MeCP2). Data were aligned to the hg38 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.