Drosophila Spike-In Control Kit for CUT&Tag (Mouse) #19629
- C&T
Product Information
Storage
实验步骤
Product Description
The Drosophila Spike-In Control Kit for CUT&Tag (Mouse) is conveniently designed to provide the reagents required to allow for normalization of variabilities across the entire CUT&Tag experiment using mouse primary antibodies. It starts with the addition of cryopreserved Drosophila spike-in nuclei to cell samples prior to performing the CUT&Tag assay. Like the cells of interest, the spike-in nuclei also bind to the Concanavalin A beads, providing a method to immobilize the spike-in nuclei for subsequent washes and incubations. During the antibody incubation step, an H2Av Mouse Monoclonal Antibody is added to the reaction in addition to the mouse primary antibody against the target of interest. This H2Av Mouse Monoclonal Antibody provides a reliable method to target, digest, and enrich histone H2Av-bound Drosophila chromatin in a consistent way across all samples. This enrichment of H2Av-bound Drosophila chromatin occurs in the same tube as the enrichment of the target of interest from the test cell chromatin, ensuring that the spike-in control nuclei are subjected to all of the same steps and conditions as the test cells. A normalization factor is generated based on the Drosophila chromatin enrichment signal and is applied to the test genome to normalize signals across samples (see Figures 1-3). Thus, this spike-in normalization approach enables reliable and consistent normalization across samples, correcting for variations in starting cell number, technical variation among samples during processing, global histone modification changes, and nonspecific enrichment coming from isotype control antibodies (i.e., nonspecific mouse IgG). This kit is intended for use only in combination with our CUT&Tag Assay Kit #77552 or other validated in-house CUT&Tag protocols. It includes one 165 µL vial of cryopreserved Drosophila Spike-In Nuclei Control, providing enough nuclei for 24 CUT&Tag reactions using a mouse primary antibody with 100,000–250,000 cells or 1–2.5 mg of tissue per reaction, when used at the recommended volume. Also included is the H2Av Mouse Monoclonal Antibody, which specifically detects the Drosophila histone variant H2Av, showing no cross-reactivity to mammalian histones. If CUT&Tag assays are performed with rabbit primary antibodies, DO NOT use this kit. Please refer to our Drosophila Spike-In Control Kit for CUT&Tag (Rabbit) #29811 for sample normalizations.
Specificity / Sensitivity
Background
A major challenge in comparing CUT&Tag datasets is being able to account for differences in signal resulting from sample-to-sample variability, such as differences in starting cell number, technical variations among samples during processing, and variations in global histone modifications. The Drosophila Spike-In Control Kit for CUT&Tag (Mouse) provides a complete normalization strategy that allows for normalization of variabilities across the entire experiment (4,5), ensuring a more accurate measurement of protein–DNA interactions between samples and across experiments.
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专