ANALYTICAL PROTEIN PROFILING

蛋白质分析

• 批准号: 7959153
• 负责人: MICHAEL D POWELL
• 金额: $ 16.94万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959153
• 关键词: Analytical Chemistry; Animals; Biochemical; Biological; Biological Assay; Biomedical Research; Blood capillaries; Cells; Charge; Computer Retrieval of Information on Scientific Projects Database; Computer software; Coupled; Data; Databases; Digestion; Equipment; Fee Schedules; Fees; Flame Ionization; Fluorescence; Funding; Gel; Glutathione; Grant; High Pressure Liquid Chromatography; Human; Image; Immunoassay; Institution; Label; Laboratories; Maintenance; Manufacturer Name; Mass Fragmentography; Modeling; Modification; Nitrogen; Operative Surgical Procedures; Oxidation-Reduction; Performance; Pharmaceutical Preparations; Phosphorus; Phosphorylation; Post-Translational Protein Processing; Price; Process; Proteome; Proteomics; Reader; Reagent; Research; Research Personnel; Resources; Running; Sampling; Scanning; Series; Services; Source; Spectrophotometry; Spottings; Structure; System; Technology; Testing; Time; Trypsin; Ubiquitination; United States National Institutes of Health; Work; base; capillary; cost; detector; drug testing; instrumentation; liquid chromatography mass spectrometry; mass spectrometer; nitration; selected ion monitoring; small molecule; synthetic polymer Bioplex

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Proteomics Lab  Michael Powell, Leader The main instrumentation of the Proteomics core lab consists of: 1. A ThermoFinnigan LTQ mass spectrometer coupled to an Agilent 1100 HPLC system to provide LC/MS/MS analysis. 2. A BioRad automated spot cutter (Proteome Works) to allow the differential analysis of 1D and 2D gels. 3. A BioRad FX Imager that allows the imaging of 2D DIGE gels and other fluorescently labeled gels. 4. A BioRad BioPlex 100 bead sorter, which allows the performance of multiple analyte assays using Luminex bead technology. Analytical Chemistry Lab  Daniel von Deutsch, Leader The newly renovated MSM ACL is equipped with multiple means to perform chromatographic and spectrographic analysis of biological samples. High Performance Liquid Chromatography (HPLC). The ACL has two Agilent (HP) 1100 series HPLCs. The available detectors include: 1) Diode Array Detector (DAD), 2) Variable Wavelength Detector (VWD), 3) 1046A Fluorescence Detector (FD), 4) an 1100 series Fluorescence Detector (FD). Optional detectors include a high sensitivity Gilson fluorescence detector, an Agilent 1049A Electrochemical Detector (ECD) and an ESA Coulochem II ECD system outfitted with multiple Model 5011 high sensitivity two channel analytical cells. Gas Chromatography/Mass Spectrometry (GC/MS). In addition, the ACL is equipped with two Agilent (HP) GCs for the detection of drugs and their metabolites. These include an Agilent (HP) 6890 Series capillary GC with Flame Ionization Detector (FID) and Nitrogen Phosphorus Detector (NPD) and an Agilent (HP) 5890 Series capillary GC coupled with 5972 Series Mass Spectrometer (MS - EI). Data can be acquired in the SCAN or Selected Ion Monitoring (SIM) modes. Spectrophotometry. The ACL is equipped with a Spectromax Plus is a UV/VIS variable wavelength microtiter plate reader with Softmax Pro software used for performing colorimetric spectrophotometry and immunoassays. This equipment allows the ACL to offer qualitative and quantitative analysis of small molecules from either human or animal samples. In addition to standard pharmacological or biochemical testing, the ACL is also capable of performing other tests that includes: drug identification, chiral separations and analysis, and redox analysis (determining levels/ratios of reduced and oxidized molecules such as glutathione). Service Charges: Proteomics Lab/2D Gel Lab We currently have a fee schedule for all services. For example, we charge by the LC/MS/MS run for analysis on the LTQ. This charge includes a digestion of the sample by trypsin, an LC/MS run, and the subsequent database searches. Database searches including post translational modifications incur an additional charge for typical modifications such as phosphorylation, nitration, ubiquitination, etc. Other more complicated analysis are charged from an estimate of the time required. Luminex assays are offered from any manufacturer. The charges are based on the price of the kit plus a setup fee of $200 per assay. Analytical Chemistry Lab The Analytical Chemistry Laboratory has a standard fee schedule for services. The fee structure is based on the amount of time required to process the samples (extraction, derivatization, etc) and the cost of supplies and reagents. In addition, a general fee is requested to help cover equipment maintenance and costs of operation.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蛋白质组学实验室  迈克尔·鲍威尔，领导者 蛋白质组学核心实验室的主要仪器包括：1。ThermoFinnigan LTQ质谱仪与Agilent 1100 HPLC系统偶联，以提供LC/MS/MS分析。 2. BioRad自动斑点切割器（Proteome Works），用于1D和2D凝胶的差异分析。 3. BioRad FX成像仪，允许2D DIGE凝胶和其他荧光标记凝胶成像。 4. BioRad BioRad 100微珠分选仪，可使用Luminex微珠技术进行多种分析物检测。 分析化学实验室  丹尼尔冯多伊奇，领导人 新装修的MSM ACL配备了多种手段来进行生物样品的色谱和光谱分析。 高效液相色谱法（HPLC）。ACL有两个Agilent（HP）1100系列HPLC。可用的探测器包括：1）二极管阵列检测器（DAD），2）可变波长检测器（VWD），3）1046 A荧光检测器（FD），4）1100系列荧光检测器（FD）。可选检测器包括高灵敏度Gilson荧光检测器、Agilent 1049 A电化学检测器（ECD）和配备多个5011型高灵敏度双通道分析池的ESA Coulochem II ECD系统。 气相色谱/质谱（GC/MS）。此外，ACL还配备了两个Agilent（HP）GC，用于检测药物及其代谢物。这些包括具有火焰离子化检测器（FID）和氮磷检测器（NPD）的Agilent（HP）6890系列毛细管GC和与5972系列质谱仪（MS-EI）耦合的Agilent（HP）5890系列毛细管GC。可以在扫描或选择离子监测（SIM）模式下采集数据。 分光光度法ACL配备了Spectromax Plus，是一种UV/维斯可变波长微量滴定板读数器，配有Softmax Pro软件，用于进行比色分光光度法和免疫测定。 该设备使ACL能够对人类或动物样本中的小分子进行定性和定量分析。除了标准的药理学或生化测试外，ACL还能够进行其他测试，包括：药物鉴定，手性分离和分析，以及氧化还原分析（确定还原和氧化分子（如谷胱甘肽）的水平/比率）。 服务费用： 蛋白质组学实验室/2D凝胶实验室 我们目前有一个收费表的所有服务。 例如，我们通过LC/MS/MS运行LTQ进行分析。 该费用包括胰蛋白酶消化样品、LC/MS运行和后续数据库检索。 包括翻译后修饰在内的数据库搜索会对典型的修饰（如磷酸化、硝化、泛素化等）产生额外费用。其他更复杂的分析会根据所需时间的估计收费。 Luminex检测试剂盒由任何制造商提供。 费用是基于试剂盒的价格加上每次检测200美元的设置费。 分析化学实验室 分析化学实验室有一个标准的服务收费表。费用结构是基于处理样品（提取、衍生化等）所需的时间以及用品和试剂的成本。此外，还要求支付一般费用，以帮助支付设备维修和业务费用。