bd fastimmune™ cytokine flow cytometry

... Intracellular Molecules for Flow Cytometric Analysis 61 Chapter 5: BD FastImmune™ Cytokine Flow Cytometry 85 Chapter 6: BD ELISPOT Assays for Cells ... with BD FACSComp Software and BD CaliBRITE Beads Perform instrument start up Perform flow check Prepare tubes of BD CaliBRITE Beads and open BD FACSComp Software Launch BD FACSComp Software Run BD ... Stream-in-air instruments include the BD FACStar™ Plus and BD FACSVantage™ (BD Biosciences Immunocytometry Systems, San Jose, CA) flow cytometers Reagents Provided Each BD CBA Kit includes specific Capture...

Ngày tải lên: 11/04/2014, 10:34

... and p19ARF act in overlapping pathways in cellular immortalization Nat Cell Biol 2:148–155 Chang BD, Swift ME, Shen M, Fang J, Broude EV, Roninson IB (2002) Molecular determinants of terminal growth ... cell senescence and tumour-promoting activities of stromal fibroblasts Cancer Lett 179:1–14 Rowland BD, Denissov SG, Douma S, Stunnenberg HG, Bernards R, Peeper DS (2002) E2F transcriptional repressor...

Ngày tải lên: 25/10/2013, 21:20

... factor NF-jB can be triggered by exposure of cells to a multitude of external stimuli, including the cytokines tumor necrosis factor (TNF-a) and interleukin-1a (IL-1a), thus initiating numerous and ... transcription Oncogene 25, 66856705 26 DiDonato JA, Hayakawa M, Rothwarf DM, Zandi E & Karin M (1997) A cytokine- responsive IkappaB kinase that activates the transcription factor NF-kappaB Nature 388, ... Shevchenko A, Bennett BL, Li JW, Young DB, Barbosa M, Mann M, Manning A et al (1997) IKK-1 and IKK-2: cytokineactivated I kappa B kinases essential for NF-kappa B activation Science 278, 860866 28 Zandi...

Ngày tải lên: 16/03/2014, 11:20

... previously [24] The cells were expanded on poly-D-lysine- and fibronectin-coated culture dishes (Falcon; BD Biosciences, Oxford, UK) in ‘complete medium’: DMEM (Gibco; Invitrogen Ltd, Paisley, UK) supplemented...

Ngày tải lên: 06/08/2014, 18:20

... with annexin V-FITC and PI for 15 minutes at room temperature Analysis was performed on a BD FacsCalibur flow cytometer data not shown) P50, p52 levels were unchanged with various drug concentrations ... described above Drug treatment was hrs after transfection of the 293 cells for a total of 150 hrs Flow Cytometry For cell cycle analysis, cells treated with or without drugs were collected by low ... analyzed using CELLQuest software (BD Biosciences) Detection of apoptosis through annexin V and PI staining was done according to the manufacturers protocol (BD Pharmingen, San Jose, CA) In brief,...

Ngày tải lên: 10/08/2014, 05:21

... survivors relative to the number of colony forming cells before the heat shock is plotted (c,d) Flow cytometry analysis of germinating spores Purified spores were prepared from a diploid strain ... microscope system (Applied Precision Inc., Issaquah, WA, USA) Heat shock analysis, budding index and flow cytometry analysis Materials and methods Strains of Saccharomyces cerevisiae All strains are ... the microscope at each time point, using a hemacytometer DNA content of cells was analyzed by flow cytometry (FACS) Cells were fixed in 70% ethanol, treated with RNaseA and proteinase K and then...

Ngày tải lên: 14/08/2014, 08:20

... Number 47 2.22 Colony Formation Assay 48 2.23 Cell Cycle Profile and TUNEL Staining Analyses by 48 Flow Cytometry 2.24 Cell Synchronization at the G2/M Boundary and 49 Cell Cycle Progression Analyses ... the absence or presence of decoy peptide treatment, with or without FBW7 rescue 108 FIGURE 21F Flow cytometry analysis of *Br1- or *Br2-induced sub-diploidization in the absence or presence of ... E2F-responsive and TRIP-Br coregulated gene Treatment with TRIP-Br decoy peptides is associated with subdiploidy 5.1 The TRIP-Br decoy peptides induce accumulation 89 of a sub-G1 population 5.2 Sub-diploidy...

Ngày tải lên: 16/09/2015, 17:12

... analysis 46 2.7 Subcellular fractionation 48 2.8 Fluorescence microscopy and neurite assay 48 2.9 Flow cytometry 49 2.10 siRNA interference 50 iv Chapter 3: Results 56 3.1 Cloning and characterization ... as molecular cloning, cell culture and transfection, real-time PCR, western blot analysis and flow cytometry analysis were employed in this study Cell cycle analysis was performed to look at events ... 8000 x g The flow- through was then discarded A volume of 700µl Buffer RW1 was added to the RNeasy column and centrifuged for 15 sec at more than 8,000 x g to wash the column The flow- through...

Ngày tải lên: 08/11/2015, 17:09

... thy-noc experiments are quite inconsistent with the proposal that the cells are synchronized The flow cytometric data on DNA contents during growth of the arrested cells shows the cells are not...

Ngày tải lên: 13/08/2014, 23:20

... Hartwell and colleagues proposed that a simple order of dependent events (e.g budding, DNA synthesis, cytokinesis), completion of which were necessary for cell division In this way, the many of the ... inactivation of E2Fs Combination of E2f 1-, or E2f 3-deficiency rescues the mid-gestational lethality of Rbdeficiency and reduces the penetrance of neuroendocrine tumorigenesis in Rb+/- mice (Tsai et al ... neurogenesis and haematopoiesis Nature 359:288–294 Lee EY, Cam H, Ziebold U, Rayman JB, Lees JA, Dynlacht BD (2002) E2F4 loss suppresses tumorigenesis in Rb mutant mice Cancer Cell 2:463–472 252 L Yamasaki...

Ngày tải lên: 25/10/2013, 21:20

... areas) Within the pocket domain (pocket subdomains A and B and the spacer region) the highest homology between the pRb protein family is found in the A and B subdomains The pocket-domain is responsible ... cellular proteins Although the binding site for LXCXE motif containing proteins is present in the B subdomain, the crystal structure of the pRb A/B pocket bound to the LXCXEcontaining part of HPV-16 ... 2001; Farkas et al 2002; Litovchick et al 2004) In p130 the Loop region, a part of the B-pocket subdomain, which is not shared with pRb nor p107, is in particular subject to phosphorylation by GSK3β...

Ngày tải lên: 25/10/2013, 21:20

... rio1-deletion strain was rescued by plasmid pMA221 which carried a RIO1 wild-type copy in the vector pGBDU-C1 [37] The coding regions of the (S > A)6 or (S > D)6 mutant alleles were amplified by PCR YMA69...

Ngày tải lên: 18/02/2014, 16:20

Ngày tải lên: 06/03/2014, 08:20

... transferred to a 30% (v/v) sucrose solution in NaCl/Pi for 24 h and embedded in OCTTM 11 compound (BDH) Immunohistochemistry was performed on consecutive fresh-frozen sections that were postfixed...

Ngày tải lên: 07/03/2014, 16:20

... the conditions used; otherwise, the immunochemical responses of the antibodies were specific Lambda protein phosphatase (400 000 unitsÆmL)1; > 300 000 UÆmg protein)1) was from Calbiochem All other ... dephosphorylate histone H1, extracts were incubated for 30 at 30 °C in the presence of 800 U Lambda protein phosphatase, 50 mM Tris/HCl pH 7.5, mM dithiothreitol, mM MnCl2, and 100 lgÆmL)1 BSA ... macroplasmodia (C) ELISA of histone H1 in cytoplasmic extracts without (m) and after incubation with Lambda protein phosphatase (j); one relative unit ¼ 105 · A490/A595 (A490 ELISA readings, A595 protein...

Ngày tải lên: 16/03/2014, 18:20

... phosphatase buffer (100 mM NaCl; 0.1 mM MnCl2; 0.1 mM EGTA; 50 mM Tris/HCl, pH 7.5) Treatment with lambda protein phosphatase (400 U; New England BioLabs) was in 0.05 mL buffer for 30 on ice and 30 ... (Cy) and of nuclear extracts (450 mM NaCl) were incubated with buffer only or with buffer plus lambda phosphatase as indicated The proteins were analysed by Western blotting (A) and autoradiography...

Ngày tải lên: 17/03/2014, 23:20

... average DNA fragment length of 0.2–0.5 kb and subjected to ChIP assay as described previously [12] Flow cytometry analysis RNA purification and RT-PCR For RNA purification, cells were resuspended in ... analyzed by flow cytommetry, using a Fusing Beckman Coulter (Fullerton, CA, USA) Cytomics FC500 Flow Cytometry Analyzer Goldberg AD, Allis CD & Bernstein E (2007) Epigenetics: a landscape takes ... HeLa cells were treated with MLL1 and scramble antisense separately for 24 h, and subjected to flow cytometry analysis (A) Control cells treated with no antisense (B) Cells treated with phosphorothioate...

Ngày tải lên: 23/03/2014, 06:20

... 60, 90, 120 and 180 (A) The cell cycle distribution of each cell population was determined by flow cytometry (B) At different times of release cells were incubated with DRB and mRNA decay was determined...

Ngày tải lên: 30/03/2014, 20:20

... were double-stained with FITC-labelled anti-BrdU and propidium-iodide (PI), and analysed by flow cytometry Fig HR12 suppresses in vitro monolayer ‘wound healing’ of Rat1/ras cells Rat1/ras cells...

Ngày tải lên: 31/03/2014, 01:20

... additional role for MEN in cytokinesis has also been identified Mob1, a MEN component, relocalizes from the SPB to the bud neck in late mitosis, where it functions in cytokinesis (176) Such relocalization ... keep it out of the nucleolus until cytokinesis is complete (186,187) The molecular basis of the relationships between mitotic exit and both the spindle and cytokinesis checkpoints are being actively ... of cytokinesis J Cell Biol 146, 777–790 185 Guertin, D A., Chang, L., Irshad, F., Gould, K L., and McCollum, D (2000) The role of 185 the sid1p kinase and cdc14p in regulating the onset of cytokinesis...

Ngày tải lên: 08/04/2014, 12:50