Disuccinimidyl Suberate is a water-insoluble, homobifunctional N-hydroxysuccinimide ester (NHS ester), Disuccinimidyl Suberate does not possess a charged group and is lipophilic and membrane-permeable, which makes it useful for intracellular and intramembrane conjugations.
NHS esters react efficiently with primary amino groups (-NH2) in pH 7 to 9 buffers to form stable amide bonds. The reaction results in the release of N-hydroxysuccinimide. Proteins, including antibodies, generally have several primary amines in the side chain of lysine (K) residues and the N-terminus of each polypeptide that are available as targets for NHS-ester reagents. Disuccinimidyl Suberate is hydrophobic and dissolved in an organic solvent such as DMSO or DMF then added to the aqueous reaction mixture.
Application:
1. Chemical cross-linking of intracellular proteins prior to cell lysis and immunoprecipitation.
2. “Fix” protein interactions to identify weak or transient protein interactions.
3. Protein cross-linking generates bioconjugates through a single-step reaction.
4. Immobilize proteins to amine-coated surfaces.
References:
[1].Cox G W, Mathieson B J, Giardina S L, et al. Characterization of IL-2 receptor expression and function on murine macrophages[J]. Journal of immunology (Baltimore, Md.: 1950), 1990, 145(6): 1719-1726.
[2].Knoller S, Shpungin S, Pick E. The membrane-associated component of the amphiphile-activated, cytosol-dependent superoxide-forming NADPH oxidase of macrophages is identical to cytochrome b559[J]. Journal of Biological Chemistry, 1991, 266(5): 2795-2804.
[3].Partis M D, Griffiths D G, Roberts G C, et al. Cross-linking of protein by ω-maleimido alkanoyl N-hydroxysuccinimido esters[J]. Journal of Protein Chemistry, 1983, 2: 263-277.
Disuccinimidyl Suberate是不溶于水的同型双功能的N-羟基琥珀酰亚胺酯(NHS-酯)。Disuccinimidyl Suberate不具有带电荷的基团，所以是亲脂性和膜可渗透的，能够用于细胞内和膜内蛋白质的共轭反应。
NHS-酯能够与pH7～9缓冲液中的伯氨基(-NH2)有效反应形成稳定的酰胺键，释放N-羟基琥珀酰亚胺。包括抗体在内的蛋白质通常在赖氨酸(K)残基的侧链中具有几个伯胺，并且每种多肽的N末端也可用作NHS-酯试剂的靶标。Disuccinimidyl Suberate是疏水性的，可先溶解在有机溶剂如DMSO或DMF中，然后再加入到水溶剂反应混合物中。
Disuccinimidyl Suberate 实际应用：
1.细胞裂解和免疫沉淀前细胞内蛋白的化学交联；
2.“固定”蛋白相互作用，以便识别较弱或瞬时的蛋白相互作用；
3.蛋白交联通过单步反应生成生物偶联物；
4.将蛋白固定到胺包被的表面上。