样品分析方案
简要概述
准备测试样品和空白样品（50 µL）
根据需要用抑制剂处理样品
加入Mca-APK（Dnp）ACE2底物工作溶液（50 µL）
在37°C下孵育样品
在Ex / Em = 320/430 nm（截止= 420 nm）下检测荧光
储备溶液配制
Mca-APK（Dnp）ACE2底物原液
用适量的DMSO准备5-10 mM的储备溶液。
注意：将溶液分装后在-20°C下，避光保存。
工作溶液配制
Mca-APK（Dnp）ACE2底物工作溶液
在1 mL分析缓冲液中加入5 µL Mca-APK（Dnp）ACE2底物储备液（10 mM），制成Mca-APK（Dnp）ACE2底物工作液。
注意：应根据经验测量底物的适当浓度。
注意：最终浓度为50 µM。
注意：100 mM含1 M NaCl，pH约7.5的Tris-HCl可用作检测缓冲液。也可以将600 µM的ZnCl2添加到缓冲液中。
操作步骤
以下方案可用作参考。
- 准备并添加50 µL测试样品和空白对照。
- 向样品和空白样品中加入50 µL Mca-APK（Dnp）ACE2底物工作溶液。
- 在37°C下孵育样品。
- 用荧光酶标仪在Ex / Em = 320/430 nm（截止= 420 nm）下检测荧光增加。
对于动力学读数：立即开始连续不断地测量荧光强度，并每5分钟记录一次数据，持续30-120分钟。
对于终点读数：在避光的条件下，将反应在所需温度下孵育30至120分钟。然后检测荧光强度
参考文献
A Comprehensive Workflow for Applying Single-Cell Clustering and Pseudotime Analysis to Flow Cytometry Data.
Authors: Melsen, Janine E and van Ostaijen-Ten Dam, Monique M and Lankester, Arjan C and Schilham, Marco W and van den Akker, Erik B
Journal: Journal of immunology (Baltimore, Md. : 1950) (2020)
Acquisition of High-Quality Spectral Flow Cytometry Data.
Authors: Fox, Amy and Dutt, Taru S and Karger, Burton and Obregón-Henao, Andrés and Anderson, G Brooke and Henao-Tamayo, Marcela
Journal: Current protocols in cytometry (2020): e74
HTLV infected individuals have increased B-cell activation and proinflammatory regulatory T-cells.
Authors: Kjerulff, Bertram and Petersen, Mikkel Steen and Rodrigues, Candida Medina and da Silva Té, David and Christiansen, Mette and Erikstrup, Christian and Hønge, Bo Langhoff
Journal: Immunobiology (2020): 151878
High-Dimensional Data Analysis Algorithms Yield Comparable Results for Mass Cytometry and Spectral Flow Cytometry Data.
Authors: Ferrer-Font, Laura and Mayer, Johannes U and Old, Samuel and Hermans, Ian F and Irish, Jonathan and Price, Kylie M
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
High-dimensional analysis of intestinal immune cells during helminth infection.
Authors: Ferrer-Font, Laura and Mehta, Palak and Harmos, Phoebe and Schmidt, Alfonso J and Chappell, Sally and Price, Kylie M and Hermans, Ian F and Ronchese, Franca and le Gros, Graham and Mayer, Johannes U
Journal: eLife (2020)
Multispectral Flow Cytometry: Unaddressed Issues and Recommendations for Improvement.
Authors: Parks, David R
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Panel Design and Optimization for High-Dimensional Immunophenotyping Assays Using Spectral Flow Cytometry.
Authors: Ferrer-Font, Laura and Pellefigues, Christophe and Mayer, Johannes U and Small, Sam J and Jaimes, Maria C and Price, Kylie M
Journal: Current protocols in cytometry (2020): e70
Phenotypic Analysis of the Mouse Hematopoietic Hierarchy Using Spectral Cytometry: From Stem Cell Subsets to Early Progenitor Compartments.
Authors: Solomon, Michael and DeLay, Monica and Reynaud, Damien
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2020)
Spectral flow cytometry-Quo vadimus?
Authors: Robinson, J Paul
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2019): 823-824
Time-Delayed Integration-Spectral Flow Cytometer (TDI-SFC) for Low-Abundance-Cell Immunophenotyping.
Authors: Hu, Wenting and Soper, Steven A and Jackson, J Matt
Journal: Analytical chemistry (2019): 4656-4664
参考文献
Analysis of the susceptibility to COVID-19 in pregnancy and recommendations on potential drug screening.
Authors: Zhao, Xiaoxuan and Jiang, Yuepeng and Zhao, Yang and Xi, Hongyan and Liu, Chang and Qu, Fan and Feng, Xiaoling
Journal: European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology (2020): 1209-1220
Angiotensin converting enzyme 2 and angiotensin (1-7) axis in pulmonary arterial hypertension.
Authors: Sandoval, Julio and Del Valle-Mondragón, Leonardo and Masso, Felipe and Zayas, Nayeli and Pulido, Tomás and Teijeiro, Ricardo and Gonzalez-Pacheco, Hector and Olmedo-Ocampo, Rossana and Sisniega, Carlos and Paez-Arenas, Araceli and Pastelin-Hernandez, Gustavo and Gomez-Arroyo, Jose and Voelkel, Norbert F
Journal: The European respiratory journal (2020)
Angiotensin-[1-7] attenuates kidney injury in experimental Alport syndrome.
Authors: Choi, Hong Sang and Kim, In Jin and Kim, Chang Seong and Ma, Seong Kwon and Scholey, James W and Kim, Soo Wan and Bae, Eun Hui
Journal: Scientific reports (2020): 4225
Anti-Inflammatory Properties of Drugs Used to Control COVID-19 and their Effects on the Renin-Angiotensin System and Angiotensin-Converting Enzyme-2.
Authors: Gilzad-Kohan, Hamed and Jamali, Fakhreddin
Journal: Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques (2020): 259-277
COVID-19 and NSAIDS: A Narrative Review of Knowns and Unknowns.
Authors: Pergolizzi, Joseph V and Varrassi, Giustino and Magnusson, Peter and LeQuang, Jo Ann and Paladini, Antonella and Taylor, Robert and Wollmuth, Charles and Breve, Frank and Christo, Paul
Journal: Pain and therapy (2020)
COVID-19, coronavirus, SARS-CoV-2 and the small bowel.
Authors: Mönkemüller, Klaus and Fry, Lucia and Rickes, Steffen
Journal: Revista espanola de enfermedades digestivas : organo oficial de la Sociedad Espanola de Patologia Digestiva (2020): 383-388
Does COVID19 Infect the Brain? If So, Smokers Might Be at a Higher Risk.
Authors: Kabbani, Nadine and Olds, James L
Journal: Molecular pharmacology (2020): 351-353
Downregulation of spinal angiotensin converting enzyme 2 is involved in neuropathic pain associated with type 2 diabetes mellitus in mice.
Authors: Yamagata, Ryota and Nemoto, Wataru and Nakagawasai, Osamu and Takahashi, Kohei and Tan-No, Koichi
Journal: Biochemical pharmacology (2020): 113825
FDA-approved thiol-reacting drugs that potentially bind into the SARS-CoV-2 main protease, essential for viral replication.
Authors: Lobo-Galo, Naún and Terrazas-López, Manuel and Martínez-Martínez, Alejandro and Díaz-Sánchez, Ángel Gabriel
Journal: Journal of biomolecular structure & dynamics (2020): 1-9
Hypertension and related diseases in the era of COVID-19: a report from the Japanese Society of Hypertension Task Force on COVID-19.
Authors: Shibata, Shigeru and Arima, Hisatomi and Asayama, Kei and Hoshide, Satoshi and Ichihara, Atsuhiro and Ishimitsu, Toshihiko and Kario, Kazuomi and Kishi, Takuya and Mogi, Masaki and Nishiyama, Akira and Ohishi, Mitsuru and Ohkubo, Takayoshi and Tamura, Kouichi and Tanaka, Masami and Yamamoto, Eiichiro and Yamamoto, Koichi and Itoh, Hiroshi
Journal: Hypertension research : official journal of the Japanese Society of Hypertension (2020)
