高敏免疫沉淀试剂盒 货号:A-P-2027
ChromaFlash High Sensitivity ChIP Kit
背景资料:Protein-DNA interaction plays a critical role for cellular functions such as signal transduction, gene transcription, chromosome segregation, DNA replication and recombination, and epigenetic silencing. Identifying the genetic targets of DNA binding proteins and knowing the mechanisms of protein-DNA interaction is important for understanding cellular processes. Chromatin immunoprecipitation (ChIP) offers an advantageous tool for studying such protein-DNA interactions. It allows for the detection of a specific protein bound to a specific gene sequence in living cells using PCR (ChIP-PCR), microarrays (ChIP-chip), or sequencing (ChIP-seq). For example, measurement of the amount of methylated histone H3 at lysine 9 (meH3-K9) associated with a specific gene promoter region under various conditions can be achieved through a ChIP-PCR assay, while the recruitment of methylated H3-K9 to the promoters on a genome-wide scale can be detected by ChIP-on-chip or ChIP-sequencing. ChIP analysis requires that ChIPed DNA contains minimal background in order to reliably identify true TF-enriched regions. High background in ChIP is mainly caused ineffective wash buffers, insufficient cross-link reversal, inappropriate DNA fragment length, and residual RNA interference. To effectively capture TF/DNA complexes, which are often in low abundance, an ideal ChIP method requires having maximum sensitivity with minimized background levels. This method should also be able to enrich highly abundant protein/DNA complexes using a small amount of cells or tissues in a high throughput format. Epigentek’s ChromaFlash™ High-Sensitivity ChIP Kit is designed to achieve these goals by maximizing sensitivity and minimizing non-specific background signals.
产品描述:This kit includes a positive control antibody (RNA polymerase II), a negative control non-immune IgG, and GAPDH primers that can be used as a positive control to demonstrate the efficacy of the kit reagents and protocol. RNA polymerase II is considered to be enriched in the GAPDH gene promoter that is expected to be undergoing transcription in most growing mammalian cells and can be immunoprecipitated by a RNA polymerase II antibody but not by non-immune IgG. Immunoprecipitated DNA is then cleaned, released, and eluted. Eluted DNA can be used for various downstream applications such as ChIP-PCR, ChIP-on-chip, and ChIP-seq。
产品特点:
1·The highly efficient enrichment ratio of positive to negative control is > 500. An extremely low number of cells (as low as 2,000 cells per ChIP reaction) can be used for enriching highly abundant protein/DNA complexes;
2·Optimized buffers and protocol allow minimal ChIP background by overcoming the weaknesses that cause non-specific enrichment, thereby increasing sensitivity and specificity of the ChIP reaction;
3·Increased antibody selectivity and capture efficiency through the use of unique chimeric proteins containing the maximum number of IgG binding domains coated on the strip-wells. This allows strong binding of any IgG subtype antibodies within a wide pH range regardless if they are in monoclonal or polyclonal form;
4.Fast 5 hour procedure, from input chromatin to ready-to-use DNA eluate;
5.96-well plate format makes the assay flexible. Either (a) manual with one single reaction each time; or (b) high throughput with 24-48 reactions each time;
6.Wide downstream analysis compatibility. Compatible with various downstream analysis workflows including ChIP-PCR, ChIP-on-chip, and ChIP-seq。
保存建议:在接收到艾德寄出的试剂盒后请按照说明书建议,使用不同的保存条件或温度来保存试剂盒内组分
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