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    Thermo Fisher ez link amine peg3 biotin
    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with <t>amine-PEG3-biotin</t> (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.
    Ez Link Amine Peg3 Biotin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ez link amine peg3 biotin/product/Thermo Fisher
    Average 79 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    ez link amine peg3 biotin - by Bioz Stars, 2020-01
    79/100 stars
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    Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.

    Journal: Nucleic Acids Research

    Article Title: PAtCh-Cap: input strategy for improving analysis of ChIP-exo data sets and beyond

    doi: 10.1093/nar/gkw741

    Figure Lengend Snippet: Schematic overview for the protein attached chromatin capture (PAtCh-Cap) method. Streptavidin coated magnetic beads are first conjugated with amine-PEG3-biotin (where PEG is polyethylene glycol). After standard cross-linking, chromatin isolation and DNA shearing procedures, 10% of the sample volume is removed and incubated with the pre-conjugated beads in the presence of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride). EDC reacts with protein carboxylate groups, forming unstable o -acylisourea activated esters that can undergo nucleophilic attack by the primary amines on the amine-PEG3-biotin prosthetics. This forms a covalent amide linkage with proteins in the chromatin complexes and releases an isourea by-product. Once bead-bound, these chromatin complexes can be subjected to additional chemical processing steps in parallel with bead-bound chromatin that was separately immunoprecipitated for the target of interest.

    Article Snippet: Following two series of washes with 0.01 M PBS (pH 7.4), 50 μg of M-280 streptavidin coated Dynabeads (Thermo Fisher Scientific; equivalent to the number of beads utilized per reaction in the Active Motif ChIP-exo Kit) were conjugated with 10 μl of 50 nM EZ-link amine-PEG3-Biotin (Thermo Fisher Scientific; PEG = polyethylene glycol) in 0.01 M PBS (pH 7.4) at room temperature for 20 min.

    Techniques: Magnetic Beads, Isolation, Incubation, Immunoprecipitation