Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag Workflows

Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag is affinity purified and specific for rabbit immunoglobulins. Minimal cross reactivity with mouse, rat, human, bovine, guinea pig, and donkey IgG is observed. Use of a secondary antibody in CUT&Tag aids in signal amplification by increasing bound IgG per target epitope [Kaya-Okur, 2019]. For best results, use with CUTANA pAG-Tn5 (EpiCypher 15-1017 / 15-1117).

*Mixed Monoclonal: a pool of multiple recombinant monoclonal antibodies.

CUT&Tag was performed on 100k K562 nuclei with 0.5 µg of either IgG (EpiCypher 13-0042), H3K4me3 (EpiCypher 13-0041), or H3K27me3 (ThermoFisher MA5-11198) antibodies, followed by 0.5 µg of anti-rabbit secondary antibody for CUTANA™ CUT&Tag. The EpiCypher Direct-to-PCR CUT&Tag protocol was used. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 5.0 million reads (IgG), 6.2 million reads (H3K4me3), and 7.2 million reads (H3K27me3). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.

(Click to enlarge)

Figure 1: CUT&Tag data
CUT&Tag was performed as described above. Gene browser shots generated using the Integrative Genomics Viewer (IGV, Broad Institute) show a representative 474 kb window centered at the LAMC3 gene. The genomic distribution pattern was consistent with that expected for H3K4me3 and H3K27me3.