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北京時(shí)間10月5日17時(shí)45分,瑞典皇家科學(xué)院宣布將2022年諾貝爾化學(xué)獎(jiǎng)授予美國(guó)化學(xué)家卡羅林·貝爾托西(Carolyn R. Bertozzi)、丹麥化學(xué)家摩頓·梅爾達(dá)爾Morten Meldal以及美國(guó)化學(xué)家巴瑞·夏普萊斯(K. Barry Sharpless),以表彰他們?cè)邳c(diǎn)擊化學(xué)和正交化學(xué)研究方面的貢獻(xiàn)。繼F.Sanger之后,Sharpless先生也兩度成為諾貝爾化學(xué)獎(jiǎng)的得主。2001年,諾貝爾獎(jiǎng)的一半授予了Sharpless以表彰他在不對(duì)稱(chēng)催化氧化領(lǐng)域的貢獻(xiàn)(另一半授予了William S. Knowles和Ryoji Noyori三位化學(xué)家以表彰他們?cè)凇?
立春,二十四節(jié)氣之首,立是開(kāi)始,春是希望。冬去春來(lái),萬(wàn)物漸次復(fù)蘇。如期而至的不止是春天,還有疫情過(guò)后的平安喜樂(lè)。一年之計(jì)在于春!一元復(fù)始,萬(wàn)象更新。在每年的春天,都要做好人生的打算,選擇好努力的方向,計(jì)劃好奮斗的目標(biāo),愿你我不負(fù)春光,不負(fù)夢(mèng)想??!立春后五日,春態(tài)紛婀娜。——白居易《立春后五日》
The copper-catalyzed azide-alkyne cycloaddition reaction, or CuAAC, has been used extensively for the conjugation, immobilization, and purification of biomolecules as well as for monitoring various biological processes. Recent advances in design of copper chelating ligands such as THTPA or BTTAA improved kinetics of copper-catalyzed azide-alkyne cycloaddition reaction and greatly increased sensitivity of alkyne detection. In spite of many recent …
Figure 1). Kinetic comparison of chelating azide and non-chelating conventional azideRecent advances in the design of copper-chelating ligands, such as THPTA or BTTAA that stabilize the Cu(I) oxidation state in aqueous solution, improve the kinetics of the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction and greatly increase the sensitivity of alkyne detection. Copper-chelating ligands have also been shown to increase the biocompatibi…
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Click-&-Go IsoTAG Kit for Profiling Intact GlycopeptidesWhile there has been much interest in profiling the intact glycoproteome, the complexity of glycoproteoforms (and more broadly, all proteoforms) remains challenging to completely define. Mass spectrometry (MS) is commonly employed for characterization of complex proteomic samples. A popular strategy for protein identification is the bottom-up shotgun proteomics approach. In this method, …
Metabolic Labeling of Glycoproteins 下載為PDFGeneral Information This protocol details the preparation of cell lysates metabolically labeled with azido- or alkynylsugar. However, investigators may wish to determine the optimal concentration of metabolic reagent as well as labeling time individually for metabolic labeling for…
Metabolic Labeling with AHA or HPG 下載為PDFGeneral Information For imaging, culture cells on cover slips or other desired imaging-compatible surface, approximately 1 × 106 cells are sufficient for one detection. 1 × 106 cells are required to prepare 100 μg of lysate. Metabolic Labeling of Cells The following is a standa…
Labeling cells with EdU 下載為PDFThe following is a general protocol was tested with several cell lines including A549, HeLa, and NIH/3T3 cells, but it can be used for any adherent cell type. Growth medium, cell density, cell type variations, and other factors may influence labeling. We recommend 10 μM E…
EdU Imaging, Flow Cytometry 下載為PDFThis protocol was tested with A549, HeLa, and NIH/3T3 cells but it can be adapted for any adherent cell type. In initial experiments, we recommend starting with 1.5 μM final concentration of an azide detection reagent. Slight adjustment of an azide detection reagent concentration might be r…
EdU Imaging, Microscopy 下載為PDFThis protocol was tested with A549, HeLa, and NIH/3T3 cells but it can be adapted for any adherent cell type. In initial experiments, we recommend starting with 5 μM concentration of an azide detection reagent. Slight adjustment of an azide detection reagent concentration might be r…
Cell Lysate Labeling 下載為PDF This is a general protocol for labeling cell lysate through copper-catalyzed click reaction with any azide or alkyne click detection reagent. This protocol may be used as a starting point for optimization of particular click c…
TCO – Tetrazine Ligation(Figure 1). Schematic representation of a TCO–Tz ligation reaction.At low biomolecule concentrations (e.g. < 5 μM) where SPAAC falls short due to modest kinetics (e.g. 0.3 to 2.3 M–1s–1), and where copper–catalyzed alkyne–azide cycloaddition click reaction might compromise system viability, trans–cyclooctene–tetrazine (TCO–Tz) is the reaction pair of choice. This ligation chemistry is based on an inverse–dema…
Cu – Free Click Chemistry (SPAAC)(Figure 1). Schematic representation of a SPAAC ligation reaction.The strain promoted alkyne-azide cycloaddition, also termed as the Cu-free click reaction, is a bioorthogonal reaction utilizing a pair of reagents, cyclooctynes and azides that exclusively and efficiently react with each other while remain inert to naturally occurring functional groups such as amines (Figure 1). SPAAC enables labeling a wide varie…
Click Chemistry Tools(CCT)位于美國(guó)亞利桑那州斯科茨代爾(Scottsdale, AZ),是一家創(chuàng)新的生物技術(shù)研究公司,專(zhuān)注于點(diǎn)擊化學(xué)(Click Chemistry)領(lǐng)域的產(chǎn)品研發(fā)和生產(chǎn)。點(diǎn)擊化學(xué)( Click chemistry)是2001年,美國(guó)諾貝爾化學(xué)獎(jiǎng)獲得者、史格堡研究院(Skaggs institute)化學(xué)生物研究所的研究員貝瑞夏普利斯(K. Barry Sharpless)發(fā)展出一種新技術(shù),其所具有的高效和高控制性。點(diǎn)擊化學(xué)的基本思想就是利用碳-雜原子成鍵反應(yīng)快速實(shí)現(xiàn)分子多樣性,一般由疊氮化物(azide)和炔烴(alk…