染色样本分析

操作步骤

1. 在DMSO中以约10 mg / mL的浓度重建Buccutite™MTA。
   注意：将未使用的MTA存放在-20°C；它最多可用于两个冷冻和解冻循环。
2. 在pH = 8.5-9.0缓冲液中以1 mg / ml以上的浓度制备目标抗体（Ab）。
3. 将MTA以8-10 µg MTA / 100 µg Ab的比例添加到Ab溶液中。
4. 充分混合并在室温下反应60分钟，在反应过程中旋转。
5. 用脱盐柱纯化反应混合物，以除去任何未反应的MTA。将缓冲液更换为PBS或您选择的其他缓冲液。
6. 收集MTA活化的抗体。通过原始起始量的70％产率估算浓度。 

1. 将预活化的PE-iFluor™660串联溶液以100 µL ddH₂O的浓度重新配制至10 mg / mL。
   注意：重建的预活化PE-iFluor™660 Tandem不稳定，不能保存超过一个月。
2. 以300µg PE-iFluor™660 Tandem / 100 µg MTA活化的Ab的比例直接将预激活的PE-iFluor™660 串联染料添加到MTA活化的目标Ab溶液中。
3. 在室温下将混合物旋转1-2小时。
4. Ab / PE–iFluor™660串联缀合物现已准备就绪。
   注意：抗体结合物应在载体蛋白（例如0.1％牛血清白蛋白）和0.02-0.05％叠氮化钠存在的情况下以> 0.5 mg / mL的浓度保存。
   注意：Ab / PE-iFluor™660串联可在4°C下保存两个月。
5. 可选：Ab / PE-iFluor™660串联可通过尺寸排阻色谱法进一步纯化以获得更好的性能。 

图示

图1.我们预活化的PE-iFluor™660串联染料已通过我们的Buccutite™FOL（提供）进行了预改性。您的抗体（或其他蛋白质）已通过我们的Buccutite™MTA（作为免费样品提供）进行了修饰，以提供经MTA修饰的蛋白质（例如抗体）。MTA修饰的蛋白质易于与FOL修饰的PE-iFluor™660串联抗体（提供）反应，以比SMCC化学方法高得多的产率得到所需的PE-iFluor™660串联抗体结合物。此外，我们的预活化PE-iFluor™660串联反应与MTA改性的生物聚合物反应的浓度远低于SMCC化学反应。

参考文献

A monolithic nanostructured-perovskite/silicon tandem solar cell: feasibility of light management through geometry and materials selection.
Authors: Elshorbagy, Mahmoud H and López-Fraguas, Eduardo and Chaudhry, Fateh A and Sánchez-Pena, José Manuel and Vergaz, Ricardo and García-Cámara, Braulio
Journal: Scientific reports (2020): 2271

Analysis of Phospholipids, Lysophospholipids, and Their Linked Fatty Acyl Chains in Yellow Lupin Seeds (Lupinus luteus L.) by Liquid Chromatography and Tandem Mass Spectrometry.
Authors: Calvano, Cosima Damiana and Bianco, Mariachiara and Ventura, Giovanni and Losito, Ilario and Palmisano, Francesco and Cataldi, Tommaso R I
Journal: Molecules (Basel, Switzerland) (2020)

Detection of Phytoestrogen Metabolites in Breastfed Infants’ Urine and the Corresponding Breast Milk by Liquid Chromatography-Tandem Mass Spectrometry.
Authors: Min, Jialing and Wang, Zitong and Liang, Chenglin and Li, Wenjie and Shao, Jie and Zhu, Kunrui and Zhou, Lu and Cheng, Jing and Luo, Shihong and Yu, Lehan and Wu, Yudong and Xie, Mingyong and Hu, Xiaojuan
Journal: Journal of agricultural and food chemistry (2020): 3485-3494

Identification and quantification of phospholipids in strawberry seeds and pulp (Fragaria × ananassa cv San Andreas) by liquid chromatography with electrospray ionization and tandem mass spectrometry.
Authors: Bianco, Mariachiara and Calvano, Cosima D and Huseynli, Lachin and Ventura, Giovanni and Losito, Ilario and Cataldi, Tommaso R I
Journal: Journal of mass spectrometry : JMS (2020): e4523

Lipid alterations in the skeletal muscle tissues of mice after weight regain by feeding a high-fat diet using nanoflow ultrahigh performance liquid chromatography-tandem mass spectrometry.
Authors: Eum, Jung Yong and Lee, Gwang Bin and Yi, Sun Shin and Kim, Il Yong and Seong, Je Kyung and Moon, Myeong Hee
Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2020): 122022

One-Pot Fabrication of Pd Nanoparticles@Covalent-Organic-Framework-Derived Hollow Polyamine Spheres as a Synergistic Catalyst for Tandem Catalysis.
Authors: Yang, Xinyi and He, Yajun and Li, Liuyi and Shen, Jinni and Huang, Jianhui and Li, Lingyun and Zhuang, Zanyong and Bi, Jinhong and Yu, Yan
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2020): 1864-1870

Supercritical CO2 Extraction and Identification of Ginsenosides in Russian and North Korean Ginseng by HPLC with Tandem Mass Spectrometry.
Authors: Razgonova, Mayya and Zakharenko, Alexander and Shin, Tai-Sun and Chung, Gyuhwa and Golokhvast, Kirill
Journal: Molecules (Basel, Switzerland) (2020)

Analysis of oxidised and glycated aminophospholipids: Complete structural characterisation by C30 liquid chromatography-high resolution tandem mass spectrometry.
Authors: Colombo, Simone and Criscuolo, Angela and Zeller, Martin and Fedorova, Maria and Domingues, M Rosário and Domingues, Pedro
Journal: Free radical biology & medicine (2019): 144-155

Evaluation of recent changes in genetic variability in Japanese thoroughbred population based on a short tandem repeat parentage panel.
Authors: Kakoi, Hironaga and Kikuchi, Mio and Tozaki, Teruaki and Hirota, Kei-Ichi and Nagata, Shun-Ichi
Journal: Animal science journal = Nihon chikusan Gakkaiho (2019): 151-157

Fast and Quantitative Phospholipidomic Analysis of SH-SY5Y Neuroblastoma Cell Cultures Using Liquid Chromatography-Tandem Mass Spectrometry and 31P Nuclear Magnetic Resonance.
Authors: Jakubec, Martin and Bariås, Espen and Kryuchkov, Fedor and Hjørnevik, Linda Veka and Halskau, Øyvind
Journal: ACS omega (2019): 21596-21603