TMR-HaloTag^® ligand

Company: Promega

Catalog#: G8251

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Verification of N-Linked Glycosylation of Proteins Isolated from Plant or Mammalian Cell Lines Using PNGase Enzyme

Wuqiang Hong, Cong Lei, Yahong Qiu, Yilan Zhou, Yili Hu, Xing Chen, Xilong Li and Jiayang Li

Published online: 2025-09-20

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Novel Protein-oligonucleotide Conjugation Method Involving a High-affinity Capture HaloTag

Author:

Junshi Yazaki,

Date:

2020-09-20

[Abstract] Highly sensitive quantitative protein profiling can play a key role in the early diagnosis of diseases, such as autoimmune diseases and cancer. We developed a modified protein-oligonucleotide conjugation method termed HaloTag-mediated barcoding, for quantifying protein molecules at a higher sensitivity than conventional protein quantification methods. This novel and efficient conjugation method can be used to prepare HaloTag-barcoded proteins using a click chemistry-based labeling technique. Here, we describe the preparation of protein-DNA complexes and detection of protein-protein interactions which can be used in a HaloTag protein barcode assay to detect an antibody. The protocol includes procedures for preparing the ligand-oligonucleotide complex, plasmid DNA preparation for protein ... [摘要] [摘要 ] 高灵敏度的定量蛋白质谱分析可以在疾病的早期诊断中发挥关键作用，例如自身免疫性疾病和癌症。我们开发了一种改良的蛋白质-寡核苷酸缀合方法，称为HaloTag 介导的条形码，用于以比常规蛋白质定量方法更高的灵敏度来定量蛋白质分子。可以使用这种基于点击化学的标记技术，将这种新颖而有效的结合方法用于制备HaloTag 条形码蛋白。在这里，我们描述了可在HaloTag中使用的蛋白质-DNA复合物的制备和蛋白质-蛋白质相互作用的检测蛋白质条形码检测以检测抗体。该方案包括制备配体-寡核苷酸复合物的程序，用于蛋白质表达的质粒DNA制备以及蛋白质-寡核苷酸复合物的制备。所描述的基于点击反应的方案简化了常规胺-酯反应方法，该方法需要色谱纯化的额外步骤。

[背景 ] 蛋白质分子可通过常规实验进行定量酶联免疫吸附测定法，western印迹和质谱的方法，例如。这些常规的定量蛋白质谱分析技术涉及使用校准曲线进行相对测量，而没有考虑DNA扩增的高灵敏度，这限制了蛋白质本身绝对量的检测。化学蛋白质组学成为可能多重测定我n中的相对定量方式，例如串联质量标签标记方法加上质谱（汤普森等人，2003 ）。蛋白质条形码技术与下一代测序技术相结合已经可以识别目标蛋白质分子。这些方法包括CITE- SEQ ，的Ab- SEQ 和L1 BRA- SEQ ...

Semi-quantitative Analysis of H4K20me1 Levels in Living Cells Using Mintbody

Author:

Yuko Sato and Hiroshi Kimura,

Date:

2017-05-20

[Abstract] Eukaryotic nuclear DNA wraps around histone proteins to form a nucleosome, a basic unit of chromatin. Posttranslational modification of histones plays an important role in gene regulation and chromosome duplication. Some modifications are quite stable to be an epigenetic memory, and others exhibit rapid turnover or fluctuate during the cell cycle. Histone H4 Lys20 monomethylation (H4K20me1) has been shown to be involved in chromosome condensation, segregation, replication and repair. H4K20 methylation is controlled through a few methyltransferases, PR-Set7/Set8, SUV420H1, and SUV420H2, and a demethylase, PHF8. In cycling cells, the level of H4K20me1 increases during G2 and M phases and decreases during G1 phase. To monitor the local concentration and global fluctuation of histone ... [摘要] 真核核DNA包裹组蛋白，形成核小体，是染色质的基本单位。组蛋白的翻译后修饰在基因调控和染色体重复中起重要作用。一些修饰是相当稳定的，作为表观遗传记忆，其他修饰在细胞周期中表现出快速更替或波动。组蛋白H4 Lys20单甲基化（H4K20me1）已显示参与染色体凝聚，分离，复制和修复。通过几种甲基转移酶PR-Set7 / Set8，SUV420H1和SUV420H2以及脱甲基酶PHF8控制H4K20甲基化。在循环细胞中，H4K20me1的水平在G2期和M期增加，G1期下降。为了监测活细胞中组蛋白修饰的局部浓度和全局波动，我们开发了一种基因编码的探针，称为薄荷素（修饰特异性细胞内抗体; Sato等人，2013和2016）。通过测量核细胞与细胞质的强度比，可以监测单个细胞中H4K20me1的相对水平。该详细方案允许甲基转移酶对基于HatoK等人的质粒H4K20me1-mintbody活性细胞中H4K20me1水平的影响进行半定量分析（2016）。

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TMR-HaloTag® ligand

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TMR-HaloTag^® ligand