Universal Magnetic Co-IP Kit
Universal Magnetic Co-IP Kit は簡便かつ迅速に共免疫沈降 (Co-IP : Co-Immunoprecipitation) を行うことができるキットです。キット付属の磁気ビーズを使用することでバックグラウンドを抑えられるとともに, 免疫沈降や磁気ビーズの洗浄操作が簡便となります。キットには, 哺乳動物細胞や組織から核抽出物および全細胞抽出物を調製するための最適の試薬およびプロトコルが含まれているため, DNA結合タンパク質複合体および細胞質に存在するタンパク質複合体どちらにも使用できます。
|Universal Magnetic Co-IP Kit Manual|
|Gene Regulation Products & Services Brochure|
|Protein-DNA Interaction Analysis Brochure|
Universal Magnetic Co-IP Kit advantages
- Perform Co-IP of nuclear or cytoplasmic protein complexes, beginning from cells or tissue
- Magnetic beads speed and simplify the procedure while reducing background, enabling the use of low-stringency buffers that maintain complex integrity
- Nuclear extraction procedure removes nuclear protein complexes from the DNA without disrupting protein/protein interactions
- Inhibitor solutions included to preserve the integrity of protein modifications
Perform Co-IP of cytoplasmic & nuclear protein complexes
While Co-IP is often used to study cytoplasmic protein complexes, traditional methods are not optimal for studying DNA-binding proteins because these nuclear complexes are fragile and can be difficult to remove from the DNA. This causes them to either be disrupted during extraction, or to be lost because they remain bound to the DNA. For this reason, in addition to containing components for whole-cell extraction, the Universal Magnetic Co-IP Kit provides nuclear extraction reagents that have been optimized to preserve nuclear protein complexes. The reduced background provided by the low non-specific binding of the kit's magnetic beads makes it possible to use a low-stringency Co-IP/Wash Buffer that helps maintain the protein complexes (Figure 1). To further improve Co-IP of nuclear complexes, the kit’s proprietary Enzymatic Shearing Cocktail uses DNA digestion to release undissociated protein complexes from the DNA. Even histone complexes, which are not released from the DNA by traditional high-salt extraction methods, can be studied (Figure 2).
Figure 1: Co-Immunoprecipitation of a nuclear complex containing SRC-1 and ERα.
Figure 2: Detection of acetylated Histone H3.
Contents & Storage
Each Universal Magnetic Co-IP Kit provides sufficient reagents to make 25 nuclear and 25 whole-cell extractions, and then to perform 25 co-immunoprecipitations. Kits include PBS, Hypotonic Buffer, Whole-cell Lysis Buffer, Co-IP/Wash Buffer, Phosphatase Inhibitors, Protease Inhibitor Cocktail, Deacetylase Inhibitor, PMSF, EDTA, Digestion Buffer, Enzymatic Shearing Cocktail, Detergent, Protein G Magnetic Beads and a Bar Magnet for making a magnetic stand. The reagent storage conditions vary from room temperature to -20°C. Please download a manual for complete kit specifications and storage conditions. All reagents are guaranteed stable for 6 months when stored properly.
The following publications cite the use of and/or provide additional information about the Universal Magnetic Co-IP Kit:
- “PTEN regulates glioblastoma oncogenesis through chromatin-associated complexes of DAXX and histone H3.3.” by Benitez et al. (2017) Nature Communications 8(15223).