saha inhibitor manufacturer br Specification table br Value

Specification table
Value of the data
Data Data on comparative antimycobacterial activity of total hydroethanolic extracts derived from five medicinal plant species is presented (Table 1). Data on in vitro cytotoxic activity of tested plant species and positive control drugs, isoniazid, rifampicin and ethambutol against MRC-5, human fetal lung saha inhibitor manufacturer cell line (ATCC® CCL-171™) is shared in Table 2.
Experimental design, materials and methods
Conflict of interest declaration
Acknowledgments Authors are thankful to the Bill and Melinda Gates Foundation (BMGF, Grant No. OPP52155) for funding the study through Noguchi Memorial Institute for Medical Research postdoctoral program on infectious diseases. We also acknowledge Mr. Heron Blogogyee of Centre for Scientific Research into Plant Medicine (CSRPM-Mampong) for assistance in plant collection, identification and preparation.
Data The dataset represents proteins that were identified to interact with integrin-linked kinase after immunoprecipitation from a model of cardiac-specific expression, with identification via mass spectrometry. The RAW data files generated from LC-MS-MS analysis of ILK immunoprecipitations have been provided, and are deposited in the ProteomeExchange (reference ID PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD001053).
Experimental design, materials and methods Hearts from ILK-transgenic mice (with cardiac-specific over-expression of ILK wild-type (WT)) were harvested and lysates prepared in ice-cold RIPA buffer [1]. Tissue extracts were prepared with RIPA lysis buffer (150mM NaCl, 1% Nonidet P40, 05% deoxycholate, 0.1% SDS, 50mM Tris, pH 8.0 and 1mM phenylmethylsulphonyl fluoride) supplemented with 1×complete protease inhibitor cocktail (EDTA-free) (Roche Diagnostics GmbH, Penzberg, Germany). The samples were centrifuged, and the resulting protein lysate (2mg) were incubated overnight at 4°C with either 2μg of anti-ILK rabbit antibody (catalogue number: 3856, Cell Signaling Technology) or control rabbit IgG (catalogue number: 011-000-003, Jackson ImmunoResearch Lab. Inc.). The samples were than incubated for 2h at 4°C with protein A agarose beads (catalogue number: sc-2001, Santa Cruz Biotechnology) and the bound beads were washed four times with RIPA lysis buffer. The protein samples were supplemented with 1×protein sample buffer, boiled and resolved on a 6-18% acrylamide gradient SDS gel. After Coomassie Blue staining, the extra bands on the IP lane contrasting to the control lane were excised from the gel and tryptically digested for MS/MS analysis. Briefly, excised gel spots were destained and washed with ACN (50%) and AMBIC (100mM), followed by drying in a vacuum centrifuge (UNIVAPO, uniEquip, Matinsried, Germany). The dried gel pieces were digested with trypsin containing digestion buffer (0.1μg/μL trypsin, 1M CaCl2, 1M AMBIC (pH 7.4)) for 45min on ice. The excess amount of trypsin solution was replaced by the same volume of 100mM of AMBIC without trypsin and incubated overnight at 37°C. The peptides were extracted with increasing concentrations of ACN and TFA and dried by vacuum centrifugation before being analyzed by mass spectrometry. Mass spectrometry was performed using an on-line liquid chromatography-tandem mass spectrometry system. An Agilent 1100 Capillary LC system (Palo Alto, CA, USA) fitted to a LTQ ion trap mass spectrometer (Thermo Electron, San Jose, CA, USA) was used for analysis. A C18 pre-column (100mm i.d×5.0cm length) and a μLC analytical column (75mm×10cm), that also served as a μESI emitter, were used for the separation of the digested proteins. The mass spectrometer was operated in data-dependent mode, automatically cycling through acquisition of a full-scan mass spectrum and three MS/MS spectra recorded sequentially. A dynamic exclusion list time of 1.5min was used. For the reverse phase chromatography, an 80-min gradient elution from water to acetonitrile, each containing 0.1% FA and 0.02% TFA, was performed at a flow of 200nl/min. All MS/MS spectra were searched against the NCBInr protein database using MASCOT Server (v2.2; Matrix Science) [2]. The search results were analysed using the Scaffold software (Proteome Software) [3]. The RAW data files are archived with the ProteomeExchange (address above). Scaffold identified 74 proteins, without a cutoff for the minimum number of spectral counts (range 18 to 1 spectral counts), although two of the proteins identified were likely the antibody used for the immunoprecipitation. To reduce the chance of identifying a non-specific interaction (i.e. “sticky” protein), the results were culled using the CRAPome database [4]. SERCA-2a possessed the largest number of spectral counts, and was fully investigated for biological relevance [5]. Other identified proteins had been investigated as ILK binding partners, including Hsp70, and published [6].