ANALYTICAL PROTEIN PROFILING

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蛋白质组学实验室负责人迈克尔·鲍威尔 蛋白质组学核心实验室的主要仪器包括：1.一台ThermoFinnigan LTQ质谱仪与安捷伦1100高效液相色谱系统联用，以提供LC/MS/MS分析。2.BioRad自动化点切割机(Proteome Works)，可对1D和2D凝胶进行差示分析。3.一种BioRad FX成像仪，允许对2D DGE凝胶和其他荧光标记的凝胶进行成像。4.BioRad BioPlex 100微珠分选机，可使用Luminex微珠技术进行多种分析物分析。 分析化学实验室负责人Daniel von Deutsch 新翻新的MSM ACL配备了多种手段，可以对生物样本进行色谱和光谱分析。 高效液相色谱(HPLC)。该ACL具有两个安捷伦(HP)1100系列HPLC。可用的检测器包括：1)二极管阵列检测器(DAD)，2)可变波长检测器(VWD)，3)1046A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、安捷伦1049A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/质谱仪(GC/MS)。此外，ACL配备了两个安捷伦(HP)GC，用于检测药物及其代谢物。其中包括带有火焰电离检测器(FID)和氮磷检测器(NPD)的安捷伦(HP)6890系列毛细管气相色谱，以及配有5972系列质谱仪(MS-EI)的安捷伦(HP)5890系列毛细管气相色谱。可以在扫描或选定的离子监测(SIM)模式下获取数据。 分光光度测定法。ACL配备了Spectrmax Plus，这是一种带有Softmax Pro软件的UV/VIS可变波长微滴定板读取器，用于执行比色分光光度和免疫分析。 该设备允许ACL对人类或动物样本中的小分子进行定性和定量分析。除了标准的药理或生化测试外，ACL还能够执行其他测试，包括：药物鉴定、手性分离和分析以及氧化还原分析(确定还原型和氧化态分子的水平/比率，如谷胱甘肽)。 服务费： 蛋白质组学实验室/二维凝胶实验室 我们目前对所有服务都有一个收费表。例如，我们按LC/MS/MS运行对LTQ进行分析。这一费用包括用胰酶消化样品，LC/MS运行，以及随后的数据库搜索。数据库搜索，包括翻译后修饰，对于典型的修饰，如磷酸化、硝化、泛素化等，都会产生额外的费用。其他更复杂的分析是从所需时间的估计中收费的。Luminex检测由任何制造商提供。费用是根据试剂盒的价格加上每次化验200美元的设置费计算的。 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。费用结构是根据处理样品所需的时间(提取、衍生等)以及用品和试剂的费用确定的。此外，还要求支付一般费用，以帮助支付设备维护费和业务费。

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.