產品介紹:
描述鏈霉親和素-生物素相互作用的高強度使得即使是高度稀釋的目標也能高效捕獲;然而,這使得從親和樹脂中回收蛋白質變得具有挑戰(zhàn)性。從固定化親和素中洗脫生物素化蛋白質的傳統(tǒng)方法包括以下幾種:(i)通過在可能包含高濃度非共價鹽的變性緩沖液中煮沸樹脂來變性鏈霉親和素;(ii)在蛋白質與樹脂結合時進行胰蛋白酶消化;或(iii)用過量自由生物素洗脫蛋白質。這些方案可能會通過釋放非特異性結合的蛋白質和/或與標記蛋白質同時釋放天然生物素化的蛋白質而共同洗脫污染蛋白。此外,部分方法可能導致與感興趣的蛋白一起洗脫高水平的樹脂基肽段,從而進一步導致樣品污染。
DADPS(雙烷氧基二苯基硅烷)生物素疊氮探針消除了鏈霉親和素-生物素親和純化的一個主要限制。這種試劑包含一個生物素基團,通過一個包含可裂解的DADPS連接器的間隔臂與疊氮基團連接。捕獲的生物分子可以在溫和條件下(5%或10%的甲酸,0.5小時)有效釋放,并且在裂解后,標記蛋白上留下的?。?43 Da)分子碎片。這些特性使得DADPS探針在生物分子標記和蛋白組學研究中尤其具有吸引力。
英文介紹:
Description
Extraordinary strength of the streptavidin-biotin interaction allows for efficient capturing of even highly dilute targets; however, it makes recovery of proteins from affinity resins challenging. Conventional methods to elute biotinylated proteins from immobilized avidin include the following: (i) denaturation of streptavidin by boiling the resin in a denaturing buffer that may include high concentrations of chaotropic salts, (ii) trypsin digestion of proteins while they are bound to the resin, or (iii) elution of proteins with excess free biotin. These protocols can co-elute contaminant proteins by releasing nonspecifically bound proteins and/or naturally biotinylated proteins concurrently with labeled proteins. In addition, some of these methods can cause elution of high levels of resin-based peptides along with the proteins of interest, resulting in further sample contamination.
DADPS (dialkoxydiphenylsilane) Biotin Azide probes eliminate a major limitation of the streptavidin-biotin affinity purification. This reagent contains a biotin moiety linked to an azide moiety through a spacer arm containing a cleavable DADPS linker. Captured biomolecules can be efficiently released under mild conditions (5% or 10% formic acid, 0.5 h) and the small (143 Da) molecular fragment left on the labeled protein following cleavage. These features make the DADPS probe especially attractive for use in biomolecular labeling and proteomic studies.
反應切割:
產品參數(shù):clickchemistrytools現(xiàn)貨DADPS Biotin Azide貨號CCT-1330選購指南
精選文獻:
1. Wang, C., et al. (2020). Chemoproteomic Profiling of Itaconation by Bioorthogonal Probes in Inflammatory Macrophages. J Am Chem Soc., 142 (25), 10894-10898.
2. Willems, L. I., et al. (2020). Tandem Bioorthogonal Labeling Uncovers Endogenous Cotranslationally O-GlcNAc Modified Nascent Proteins. J Am Chem Soc., 142 (37), 15729-15739.
3. Wang, J., et al. (2015). Mapping sites of aspirin-induced acetylations in live cells by quantitative acid-cleavable activity-based protein profiling (QA-ABPP).Sci. Rep.. 5: 7896.
4. Jinxu, G., et al. (2012). Small Molecule Interactome Mapping by Photoaffinity Labeling Reveals Binding Site Hotspots for the NSAIDs. J. Am. Chem. Soc.,. 140: 4259-68.
5. Szychowski, J., et al. (2010). Cleavable Biotin Probes for Labeling of Biomolecules via Azide?Alkyne Cycloaddition. J. Am. Chem. Soc., 132: 18351-60.
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