isolation and characterization of cells by dielectrophoretic field flow fractionation

... condition C, E and G: Undifferentiation; D, F and H: Neuronal differentiation Nestin, Tuj-1 and NF160 were green GFAP and MAP2 were red Scale bars = 50 μm Isolation and characterization of canine ... (Figs 4E and F) Discussion Isolation and characterization of stem cells derived from various tissues and sources have been one very critical issue for stem cell therapy [12,28,33] The purpose of this ... multipotent stem cells from dogs The characterization conditions and basic settings for the application of gene delivery were also investigated Materials and Methods Cell isolation and culture Canine...

Ngày tải lên: 07/08/2014, 23:22

... and mutant cells and extent of HIV Growth Growth rates of parental and mutant cells and extent of HIV integration (A) Growth rates Parental and mutant 30-2 and 42-7 cells were seeded and growth ... x 105 Figure cells with an HIV-EGFP vector at a moi = 0.5 Infection of parental and mutant HeLa cells (30-2 and 42-7) Infection of parental and mutant HeLa cells (30-2 and 42-7) cells with an ... extent of integrated HIV-1 vector was measured by infection of cells at moi = The cells were passaged times and the quantity of stable HIV-1 DNA was measured by quantitative real time PCR Page of...

Ngày tải lên: 13/08/2014, 05:22

... (Tris-HCl) buffer of pH 8.0 Elution was done with the same buffer at a flow rate of ml/h and ml fractions were collected The void volume of the column was determined by the elution volume of Blue Dextran ... polysaccharide One milligram of the partially purified polymer contained 309.7 µg of protein, 160.2 µg of neutral sugar, 58.5 µg of hexosamine, 30.8 µg of uronic acid and 3.8 µg of phosphorus Constituent ... as follows: 500 g of soil was washed times with distilled water and suspended in 1,000 ml of water The mixture was allowed to stand for 10min and the supernatant was collected and used for the...

Ngày tải lên: 05/09/2013, 09:08

... for cDNA single strand synthesis, 600 ng of RNA, 0.5 lg of oligo(dT) primer and 10 mm of dNTPs were incubated for at 65 °C Subsequently, lL of the first strand buffer X and lL of dithiothreitol ... Nomenclature of the toxic pulchellin lectins The abbreviation P is followed by the Roman numerals I, II, III and IV and refers to each pulchellin Characterization of four pulchellin isoforms isoform ... presented by a mixture of closely related isoforms In the present study, four pulchellin isoforms were isolated, and their amino acids sequences deduced by cDNA cloning and verified by MS Half...

Ngày tải lên: 18/02/2014, 16:20

... of 50 nmÆmin)1, band-width of 1.0 nm, and a resolution of 0.1 nm in the temperature range between 10 °C and 70 °C, after preincubation for 10 at each temperature Three runs were accumulated and ... of 0.6 lM The data were fitted to Michaelis–Menten curves Squares and dotted line: data and fitting with PHI/PHO ratio of 0.5 Triangles and continuous line: data and fitting with PHI/PHO ratio of ... obtain separation of PHR and PHI All steps were performed at °C Monomers isolation by reverse-phase HPLC PHI was resuspended in 80 lL of 50% water/50% acetonitrile solution and 1% formic acid...

Ngày tải lên: 21/02/2014, 00:20

... intense band of 61 kDa (NrfA) and a band of weak intensity of 19 kDa (NrfH), confirming its hetero-oligomeric nature (Fig 1, lane 1) However, in the absence of boiling (Fig 1A, lanes and 4) high ... and 4) high molecular mass bands of approximately 110 kDa and > 200 kDa were visible, as well as a faint band at 37 kDa, suggesting the presence of dimers All of the bands stained positively for ... suggests that the bands at approximately 110 and 37 kDa correspond to strongly attached dimers of both NrfA (a2) and NrfH (b2), respectively The presence of dithiothreitol (Fig 1A, lanes and 2) did not...

Ngày tải lên: 21/02/2014, 00:20

... BLAST of peptide sequences of 21 kDa proteins in databases Tryptic peptides were puri®ed by RP-HPLC and some of them were totally or partially analyzed by Edman sequencing (ááá) and/ or by MALDI-TOF ... Chlamydomonas cells Chlamydomonas CW15 cells were grown to a cell density > ´ 106 cells per mL Cells were pelleted, resuspended in 30 mM Tris/HCl pH 7.9, and broken by two cycles of freeze-thawing ... cDNA and the Ch-Prx1 gene, respectively RNA isolations RNA for RT-PCR and for northern analyses was islolated by alternative methods In the ®rst method,  30 million cells were pelleted by centrifugation...

Ngày tải lên: 08/03/2014, 16:20

... presequence of 19 amino acids The mature protein would have a molecular mass of 41.7 kDa and a pI of 6.34 The YedO protein from E coli and the d-CDes from Arabidopsis showed an overall identity of 36% and ... supported by Western blot analysis of isolated organelles and by the localization studies using fusions with GFP (Figs and 4) In general, the localization predictions of plastidic and mitochondrial ... light ⁄ dark rhythm and under sulfate limitation Results In silico characterization and isolation of the Arabidopsis protein homologous to YedO from E coli The existence of d-CDes activity was...

Ngày tải lên: 16/03/2014, 18:20

... ampicillin and/ or 2% (w/v) glucose Solid media contained 2% (w/v) Bacto-agar Transformation of E coli was by standard procedures [24] performed using electrocompetent cells Isolation of total RNA ... possibility of disulphide bond formation genetically by replacement of residues Cys22 and Cys82 with alanine and valine, respectively, to give a cysteine minus mutant (12F-11DCys) Use of alanine and ... Import and assembly of proteins into mitochondria of mammalian cells Biochim Biophys Acta 1592, 97–105 19 Suzuki, H., Maeda, M & Mihara, K (2002) Characterization of rat TOM70 as a receptor of the...

Ngày tải lên: 17/03/2014, 10:20

... selected on the basis of a nutritional marker (URA3 gene), contained within the vector, and by the ability of cells to recombine the two DNA molecules by gap repair Transformed cells were plated ... made by random priming and the incorporation of [a-32P]dCTP Hybridization of the probe to the membrane and subsequent washes were performed as described [29] Hybridization was detected by autoradiography ... strain genuinely reflects of the sensitivity of the mutant toxin to temperature To obtain a clearer picture of the growth profiles of yeast cells expressing these RTAs, cells were plated at various...

Ngày tải lên: 23/03/2014, 07:20

... Switzerland), thermolysin, chymotrypsin or elastase (Sigma, St Louis, MO, USA)] and Cloning of the bovine MUC15 cDNA by PCR with degenerate primers Isolation of total RNA from the mammary gland of ... Expression of MUC15 in human and bovine tissues and cell types MUC15 mRNA expression was examined by PCR screening of commercial multiple tissue cDNA panels and oligo(dT) primed cDNA libraries and by ... PCR screening of bovine and human cDNA libraries showed the presence of MUC15 mRNA in human breast, bovine mammary gland, bovine lymph nodes and lungs of both species (Table 1) Of the identified...

Ngày tải lên: 24/03/2014, 04:21

... of degenerate and specific primers and by rescreening the cDNA libraries The partial clones of XHIVEP2 and -3 covered 5.9 and 3.2 kb of the respective 3¢ ends and included 3¢-UTRs and poly(A)-tails ... were annealed by heating to 90 °C an equimolar mixture of the upper and lower strands in reaction buffer and cooling slowly to ambient temperature (1 h) Sequences of the upper strand of the duplexes ... functional domains of XHIVEP1 Thus, the cloning and characterization of the XHIVEP interacting factors would be of interest and should also provide additional insight into the function of this protein...

Ngày tải lên: 30/03/2014, 13:20

... streptomycin and non-essential amino acids Vero Cells were obtained from ATCC and were maintained in MEM with Earl's salts and the addition of 10% FBS and standard concentrations of L-glutamine, ... Domains of E9 and locations of mutations responsible for drug resistance Domains of E9 and locations of mutations responsible for drug resistance The location of the putative exonuclease and polymerase ... adsorption, the virus was removed and the cells washed with PBS One ml of DMEM containing 10% FBS was added to cells Infected cells were incubated at 37°C and harvested by scraping at the following...

Ngày tải lên: 20/06/2014, 01:20

... the G and P serotype specificities of the YK-1 strain, the sequences of the genes encoding both VP7 and VP4 were determined and compared with those of representatives of established G and P serotypes ... as a reference It consisted of 667 nt with a 594-bp ORF flanked by 5' and 3' UTRs of 21 and 52 nt, respectively Segment 11 of YK-1 had 99% similarity to segment 11 of RRV In the variant 311, segment ... are known, and the NSP4 of YK-1 can be classified as Group C by comparison of the amino acid sequence (aa 131–148) of the variable portion in the VP4 binding domain of various groups of NSP4 (Figure...

Ngày tải lên: 20/06/2014, 01:20

... eht fo esahp suoeuqa reppu eht no detcelloc saw AND lairetcaB nim rof 000,6 ta degufirtnec dna nim rof sllec lairetcab gniliob yb detcartxe neht saw AND lairetcab ;retaw dellitsid elbuod dezilirets ... 817( rof evitisop ,9~4 senal ;)pb 001,1( rof evitisop ,3 dna senal ;noitcefni laccocotperts rof evitagen ,1 enal ;lortnoc evitagen ,N enal ;)pb 003 ,M4040 ( lortnoc evitisop ,P senal ;reddal AND ... enilav ,esapil rof snoitcaer evitagen dah setalosi llA nispyrtomyhc -α dna esatahpsohp enilakla rof snoitcaer evitisop dah fo setalosi 81 dna esadinimasoculg-β-lyteca -N dna esaretse rof snoitcaer...

Ngày tải lên: 07/08/2014, 18:21

... assayed in duplicates and each assay was validated by comparison with positive and negative chicken and equine control sera as well as back titration of the used virus dilutions Page of Sequence analysis ... letters showed site of H274Y and N294S substitutions (H275Y and N295S in N1 influenza numbering) cough, fever and serous nasal discharge The course of the disease was short (72 h) and responsive well ... Details 1Department of Virology, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt, 2Division of Virology, Department of Microbiology, College of Medicine and Medical Sciences,...

Ngày tải lên: 10/08/2014, 05:21

... The titers (PFU mL-1) of virus-containing cultures were determined by serial dilution Plaque assay and virus isolation Figure Schema of PBCV infection of the symbiotic algae of Paramecium *Paramecium ... than the DNAs of other BW strains That is, CvV-BW1 DNA was cut only by MssI and SwaI, while CvV-BW2, -BW3, and -BW5 DNAs were effectively cut by all six enzymes tested DNAs of CvV-BW2 and -BW5 showed ... whereas the intensity of band Z increased; with heat treatment at 100°C, only band Z was observed The sizes of proteins in bands X, Y, and Z were estimated to be 370 kDa, 105 kDa, and 50 kDa, respectively,...

Ngày tải lên: 12/08/2014, 01:21

... 5.85, and 8.90 min, respectively The products were quantified by peak area and identified by comparison with the retention times and molecular ions (m/z) of methyl halide, CH3SH, and DMS standards ... sativus HT performed the isolation and heterologous expression of the HTMT encoding gene from R sativus and characterization of the enzymatic properties of recom- Page of 10 (page number not for ... absence of halide ions This data concurs with the previous reports by Fall et al [28] and Kanda et al [29,30] of the emission of sulfur-containing gases including DMS from maize, wheat, and rice...

Ngày tải lên: 12/08/2014, 03:21

... DNAstar software package Sequence analysis based on the complete ORF2 sequence of the isolate in this study and other 10 genotype HEV isolates in China, using DNAstar software package Page of (page ... participated in the planning of the project XGH was the leader of the project WZ, SXY, QS, and TLS collected all samples and performed the RT-PCR experiments for HEV RNA WZ and SXY did the phylogenetic ... Bezanilla JC, Toth TE, Meng XJ: Identification of genotype hepatitis E virus (HEV) in serum and faecal samples from pigs in Thailand and Mexico, where genotype and HEV strains are prevalent in the respective...

Ngày tải lên: 12/08/2014, 04:21

... identification and characterization of MPs in the sputa of CF patients Patients and Methods Study patients The study was approved by, and performed in accordance with, the ethical standards of our institutional ... Identification of MPs by flow cytometry Representative dot plots and histograms of MPs from sputum from CF patients and P aeruginosa and S aureus MPs (A), P aeruginosa PAO1 (B), S aureus (C) and calibrator ... CF and PCD sputum) Statistical significance of differences between acute and stable groups of CF patients was evaluated by a two-tailed unpaired Student's t-test To compare the number of MPs and...

Ngày tải lên: 12/08/2014, 11:22