practical experiences post assay development and validation

Ngày tải lên: 12/01/2014, 22:19

... Quantification and validation of 19 genes by qPCR The expression of 19 genes in test lung cancer cell lines (black and gray) and four validation cell lines (white) was quantified by qPCR and compared ... for each gene and used as the standard sample in the subsequent assay Each standard curve consisted of six serial dilutions (1, 1/5, 1/25, 1/125, 1/625, and 1/3125) of the selected standard cDNA ... S100P, and SLCO4A1) and downregulation in PLAU The validation SQ cell line showed similar upregulation in 10 genes (AMY2A, BEX1, CDH1, HMGA1, IGSF3, INADL, ISL1, MALL, RAB25 and SLCO4A1) and downregulation...

Ngày tải lên: 15/02/2014, 04:20

... function, fn and fu are fractions of native protein for pre-transition and post- transition curves, respectively, obtained by extrapolation to K, and mn and mu are slopes of the pre-transition and post- transition ... where T0 and TF are lower and upper temperature limits of transition, respectively, and Tmax is the excess heat capacity associated with the transition The Gibbs free energy, DG ẳ DH ) TDS, and thus ... ẳ a1+a2T and yU ẳ b1+b2T are the means of y, characteristic of the native and denatured conformation, respectively They were obtained by linear regression of the pre-transition and post- transition...

Ngày tải lên: 20/02/2014, 02:21

... experience and publications in the field of applications of calorimetry and biocalorimetry in the analysis of small and large drug molecules I welcomed the invitation and I was enthusiastic to handle ... Magnetic and hyperfine characterizations were studied by vibrating sample magnetometer and Mössbauer spectrometry, respectively Fe and FeCo-based alloys 6.1 Fe and Fe-Co powders Fe and Fe50Co50 ... behaviour for both Fe and Fe50Co50 powders (Fig 4) A linear fit gives slopes of 0.65 and –0.20 for short and extended milling times, respectively, in the case of Fe; and slopes of –0.85 and –0.03, respectively,...

Ngày tải lên: 06/03/2014, 22:20

... human PKIb; the bands at 1675, 1636 and 1625 cm)1 can be assigned to extended chain structures; the bands at 1683, 1669 and 1663 cm)1 are possibly assigned to turns or bends; and the band at 1647 ... The 1652 and 1270 cm)1 bands can be assigned to a-helix in the human PKIb; the 1666 and 1247 cm)1 bands are the characteristic frequencies of the random coil structure Meanwhile, the 1682 and 1255 ... 1669, 1653, 1647 and 1636 cm)1, and two shoulders at 1663 and 1625 cm)1, which indicate that the amide I¢ mode consists of various overlapping components (Fig 4A) These component bands can be better...

Ngày tải lên: 07/03/2014, 15:20

... containing mM MgCl2 and 0.5 mM ADP, stored in the same buffer, and used for experiments during three days The concentrations of t-S1 and Nt-S1 were measured by the Bradford protein assay [32] using ... and Vi (A) or BeFx (B) before and after removal of excess reagents Curves shown by dashed lines were obtained for Nt-S1 in the presence of 0.5 mM ADP and 0.5 mM Vi (A) or 0.5 mM ADP, mM NaF and ... ability of S1 to form stable complexes S1ÆADPÆVi and S1ÆADPÆBeFx and to undergo structural changes due to formation of these complexes MATERIALS AND METHODS Proteins S1 from rabbit skeletal myosin...

Ngày tải lên: 17/03/2014, 10:20

... tensors with the simplicity and concreteness of vectors General treatments of di erential forms and EM theory include papers 2], 10], 11], 12], 13], and 14] Ingarden and Jamiolkowksi 15] is an ... that the usual order of development can be reversed by introducing Faraday's and Ampere's laws to students rst and using these to motivate Gauss's laws The Stokes theorem and the divergence theorem ... intensity ^ ^ z 1-forms E and H are dual to the vectors E and H Following Deschamps, we take the units of the electric and magnetic eld intensity 1-forms to be Volts and Amps, as shown in Table...

Ngày tải lên: 24/04/2014, 16:51

... property of the sample and the temperature an enclosure within which the experimental parameters may be controlled a computer to control data collection and processing EYP 2006 TGA and DSC Thermogravimetric ... thermocouples Separate sensors and heaters for the sample and reference sample pan reference pan inert gas vacuum Furnace: inert gas vacuum thermocouple ΔT = separate blocks for sample and reference cells ... of standard DSC Fourier Transformation analysis is used to separate the total heat flow into its two components: reversing and non-reversing heat flow increased sensitivity, resolution and the...

Ngày tải lên: 24/05/2014, 20:41

... contributed to the design of the study and carried out the proliferation assays, TUNEL assays, cell cycle assays, Western blotting and statistical analysis JC and MC contributed to the interpretation ... hours 50 μl of M NaOH was added and the absorbance was read at 405 nM (reference – 620 nM), as previously described [16] Invasion assays Invasion and migration assays were performed as previously ... invasion inserts for invasion assays and uncoated inserts for migration assays Cells were incubated for hours before dasatinib treatment to allow cells to attach and then incubated at 37°C with...

Ngày tải lên: 18/06/2014, 15:20

... and As4 fluxes, 1.0 10-5 Pa and 6.0 10-4 Pa, respectively After the GaAs buffer layer growth, about 1.5 monolayers (ML) of an InAs WL were grown at 500°C under In and As4 fluxes, 1.28 10-6 Pa and ... in a hole structure under In and As4 irradiations at 300°C and its line profiles a Shows the hole structure, b and c were images after supplying additional 0.02 ML and 0.04 ML of InAs, respectively ... applying voltage was 0.052 s, and a voltage value was increased every 0.00026 s by a step function, and then observed the surface structure transitions Results and Discussion Surface Structure...

Ngày tải lên: 21/06/2014, 08:20

... Jarvis and Walker.13 Amylose (Type III from potato, amylopectin free, from Sigma Chemical Company, St Paul, MO, USA) and amylopectin (from potato, Fluka Company, Switzerland) were used as standards ... transitions were broad and flatten with Tcxo of ~119oC for 1:1, sharpen prominently and bigger for 1:2 but became slightly smaller and broader for 1:3, with Tcxo of ~89oC and 85oC, respectively ... native rice starch during cooling in DSC experiment which may have practical significance on food quality and stability MATERIALS AND METHOD 2.1 Materials Rice starch was obtained from Sigma Chemical...

Ngày tải lên: 07/08/2014, 14:20

... assisted in the animal studies and cell culture and performed immunohistochemical stainings and digital image analysis MJM and SG participated in the design of the study and helped to draft the manuscript ... were collected, and TNF-α levels were measured by sandwich enzymelinked immunosorbent assay (ELISA) Data are representative of three independent experiments performed in triplicates and are expressed ... were collected and IL-6 levels were measured by sandwich ELISA and corrected for weight of the biopsy Data are representative of three independent experiments performed in triplicates and are expressed...

Ngày tải lên: 09/08/2014, 08:22

... performed the cartilage assays, and helped conceive, design, and coordinate the study and draft the manuscript ADR and TEC helped conceive, design, and coordinate the study and draft the manuscript ... of metalloproteinases and their inhibitors and correlate this with pro-collagenase activation and aggrecan and collagen release Materials and methods Cartilage degradation assay Bovine nasal cartilage ... readily available, and this explant culture system provides a rapid, reproducible, and reliable model system to study the mechanisms of cartilage degradation and as such has become a standard for studying...

Ngày tải lên: 09/08/2014, 08:22

... factor and activates a program of cell dissociation and motility coupled with increased protease production that has been shown to promote cellular invasion [6,7] HGF and c-Met are co-expressed and ... Apoptosis and necrosis, quantified using the acridine orange assay, in SF767 cells after treatment with radiation, MP470, or Figure (A) Apoptosis and necrosis, quantified using an acridine orange assay, ... technical laboratory support DB provided resource and technical advice BS provided resources and participated in the design and coordination All authors read and approved the final manuscript 10 11 Acknowledgements...

Ngày tải lên: 09/08/2014, 10:20

... Authors’ contributions MC and KR designed the study MC and MAE collected and inputted the data MC and KR conducted the statistical analysis MC and MAE, KR compiled the data and MC & KR drafted the ... hallux limitus and its relationship to gait efficiency J Am Podiatr Med Assoc 1986, 76 15 McPoil T, Hunt G: Evaluation and management of foot and ankle disorders: present problems and future directions ... loading of the foot and patient positioning, not solely the choice of casting technique This emphasises the need for development of standardised positioning methodologies for casting and digitisation...

Ngày tải lên: 10/08/2014, 21:24

... (pSTAT1), p-STAT3 and p-STAT5 [50] Membrane-bound and soluble forms of FLT3 ligand are both biologically active [53] FLT3 ligand plays an important role in survival, proliferation, and differentiation ... proteinuria, hypertension, myalgia, skin toxicity (hand and foot blisters) and oral hypersensitivity, and these toxicities increased in frequency and intensity with increasing doses The maximal tolerated ... ABT-869 underwent the industrial standard pre-clinical toxicology, metabolism, and pharmacology studies and the compound was deemed to be suitable to further clinical development (see below) http://www.jhoonline.org/content/2/1/33...

Ngày tải lên: 10/08/2014, 22:20

... as OC The columns and bars indicate the mean and standard deviation (S.D.) from duplicated wells b) OC formation in co-culture with osteoblast (upper) and culture with M-CSF and sRANKL (lower) ... receptor, RANK, and αβν3 integrin The importance of PI3-K for differentiation, survival and motility of OCs has been demonstrated by using the PI3K inhibitors wortmannin and LY294002 [15-22], and also ... degradation of IκB and phosophorylation of JNK and ERK1/2 induced by sRANKL On the other hand, the expression of NFATc1, which occurs in the late phase of OC differentiation and promotes terminal...

Ngày tải lên: 12/08/2014, 12:20

... 7:121 http://respiratory-research.com/content/7/1/121 the EAR 3and thesaline and Y-27632 (A) and to reduce andare(PGF2α) animals PC and airway (hyper)responsiveness inhaEffectivenessrepresent for ... [9] and after repeated allergen challenge in rats [13] and mice [17] Inflammatory cells – activated during the allergic reaction – release mediators, including prostaglandins, leukotrienes and ... to histamine and PGF2α before and after the allergen-induced EAR and LAR We demonstrate that there is a differential role of Rho-kinase in the agonistinduced airway obstructions and that inhalation...

Ngày tải lên: 12/08/2014, 16:20

... designed and implemented the DDP-based feature selection technique and the subsequent analyses on the technique, and drafted the manuscript MC supervised and SWT co-supervised the study Both MC and ... model it lies between 0.1 and 0.2 for G = 3, 5, 10 and between 0.2 and 0.3 for G = 20, 30 ∗ Indeed, the observation regarding α MP provides the link between the study in [3] and the DDP concept In ... located on the i-th row and the k-th column genes from the first and 14-th groups of marker genes, respectively, while S0.3 has only and redundant genes from the first and 14-th groups of marker...

Ngày tải lên: 12/08/2014, 17:20