CUT&Tag Assay Kit #77552

The CUT&Tag Assay Kit is designed to conveniently provide reagents needed to perform up to 24 reactions. The kit has been optimized to work in fresh or lightly fixed cells for all types of DNA-binding proteins, including histones, transcription factors, and cofactors. In addition, the kit has been optimized to work for histones in fresh or lightly fixed tissues. For analysis of transcription factors and cofactors in tissues, we recommend using the CUT&RUN Assay Kit #86652. If possible, we recommend using 100,000 cells or 1 mg of tissue per CUT&Tag reaction. If starting cell number is limited, this kit is validated to work with as few as 5,000 to 10,000 cells per reaction for histone modification targets and as few as 20,000 cells per reaction for transcription factors and cofactors. This kit is compatible with both whole cells and nuclei as starting material. We have not found that using nuclei generates a stronger signal or a higher signal-to-noise reaction compared to whole cells. A complete assay can be performed in as little as one day. The CUT&Tag Assay Kit also provides important controls to ensure a successful CUT&Tag experiment, including positive control Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit Monoclonal Antibody #9751 and negative controls Normal Rabbit IgG #2729 and Normal Mouse IgG #68860. The negative control Normal Rabbit IgG #2729 or Normal Mouse IgG #68860 is optional for the experiments, depending on the requirements of the peak calling algorithm used. This kit is compatible with downstream Next Generation sequencing (NG-seq) analysis, but not qPCR.

This kit is fully compatible with the Drosophila Spike-InControl Kit for CUT&Tag (Rabbit) #29811 and Drosophila Spike-In Control Kit for CUT&Tag (Mouse) #19629, both of which provide a comprehensive sample normalization strategy that involves spiking in cryopreserved Drosophila nuclei and a Drosophila histone H2Av antibody into each CUT&Tag sample. This spike-in normalization approach enables reliable and consistent normalization across samples by correcting for variations in starting cell number, technical variability during processing, global histone modification changes, and non-specific enrichment from isotype control antibodies (i.e. non-specific rabbit IgG). The Drosophila Spike-In Control Kit for CUT&Tag (Rabbit) #29811 should be used when performing CUT&Tag with rabbit target-specific antibodies, and the Drosophila Spike-In Control Kit for CUT&Tag (Mouse) #19629 should be used with mouse target-specific antibodies. For detailed instructions, please refer to each of the spike-in control kits.