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https://www.selleckchem.com/products/art0380.html Adherent cells are easily prepared for cell staining by growing on a suitable microscope slide, coverslip, or plastic tissue culture dish. For high-resolution studies, adherent cells should be grown on the highest available grade glass coverslips, because the controlled thickness, flatness, and good optical properties of a proper coverslip are required to produce the best images. In addition, the glass surface is compatible with all fixing and staining solutions. If many antibodies, different dilutions, or various controls are to be tested on the same cell type, plating the cells onto multiwell slides can be helpful. For low-resolution work, such as crude antigen detection, hybridoma screening, or antibody titration, cells for staining can be grown on regular tissue culture dishes.Chromatin immunoprecipitation, commonly referred to as ChIP, is a powerful technique for the evaluation of in vivo interactions of proteins with specific regions of genomic DNA. Formaldehyde is used in this technique to cross-link proteins to DNA in vivo, followed by the extraction of chromatin from cross-linked cells and tissues. Harvested chromatin is sheared and subsequently used in an immunoprecipitation incorporating antibodies specific to protein(s) of interest and thus coprecipitating and enriching the cross-linked, protein-associated DNA. The cross-linking process can be reversed, and protein-bound DNA fragments of optimal length ranging from 200 to 1000 base pairs (bp) can subsequently be purified and measured or sequenced by numerous analytical methods. In this protocol, two different fixation methods are described in detail. The first involves the standard fixation of cells and tissue by formaldehyde if the target antigen is highly abundant. The dual cross-linking procedure presented at the end includes an additional preformaldehyde cross-linking step and can be especially useful when the target protein is in low abundance or if
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