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#24054642   2013/09/23 Save this To Up

Study of extraction procedures for protein analysis in plankton samples by OFFGEL electrophoresis hyphenated with Lab-on-a-chip technology.

Extraction procedures for protein analysis from plankton samples were studied. OFFGEL electrophoresis combined with Lab-on-a-chip technology has been applied for protein analysis in plankton samples. BCR-414 (plankton) certified reference material from the European Commission was used to evaluate the protein extraction procedures. Three protein extraction procedures were studied: (1) by using Tris-HCl buffer containing a protease inhibitor cocktail, (2) urea/triton X-100 buffer extraction, and (3) using the phenol/sodium dodecyl sulphate method after different washing steps with 10% trichloroacetic acid/acetone solution and methanol. The pellet of proteins obtained was dried and then dissolved in the OFFGEL buffer. Proteins were separated according to their isoelectric points by OFFGEL electrophoresis. This separation was performed using 24 cm, pH 3-10 IPG Dry Strips. The proteins present in each liquid fraction (24 fractions) were separated according to their molecular weight using a microfluidic Lab-on-a-chip electrophoresis with the Protein 80 LabChip kit. This kit allows for the separation of proteins with a molecular weight ranging from 5 to 80 kDa. Taking into account the intensity and the number of the protein bands obtained, the protein extraction procedure using the phenol/sodium dodecyl sulphate after different wash steps with 10% trichloroacetic acid/acetone solution was selected. The developed method was applied for protein determination in a fresh marine plankton sample. The proteins found in this sample have a molecular weight ranging from 6.4 to 57.3 kDa, and the proteins with highest molecular weight were in the OFFGEL fractions with an isoelectric point ranging from 4.40 to 8.60. The concentration of proteins were calculated using external calibration with Bovine Serum Albumin, and the protein concentrations varied from 50.0 to 925.9 ng µL(-1).

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HIV 1 intergase antigen. Bone Morphogenetic Protei Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Gro g Macrophage In Mouse Macrophage Inflamma Mouse Macrophage Inflamma Mouse Macrophage Inflamma Mouse Macrophage Inflamma

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#23711730   2013/05/28 Save this To Up

A practical and novel method to extract genomic DNA from blood collection kits for plasma protein preservation.

Laboratory tests can be done on the cellular or fluid portions of the blood. The use of different blood collection tubes determines the portion of the blood that can be analyzed (whole blood, plasma or serum). Laboratories involved in studying the genetic basis of human disorders rely on anticoagulated whole blood collected in EDTA-containing vacutainer as the source of DNA for genetic / genomic analysis. Because most clinical laboratories perform biochemical, serologic and viral testing as a first step in phenotypic outcome investigation, anticoagulated blood is also collected in heparin-containing tube (plasma tube). Therefore when DNA and plasma are needed for simultaneous and parallel analyses of both genomic and proteomic data, it is customary to collect blood in both EDTA and heparin tubes. If blood could be collected in a single tube and serve as a source for both plasma and DNA, that method would be considered an advancement to existing methods. The use of the compacted blood after plasma extraction represents an alternative source for genomic DNA, thus minimizing the amount of blood samples processed and reducing the number of samples required from each patient. This would ultimately save time and resources. The BD P100 blood collection system for plasma protein preservation were created as an improved method over previous plasma or serum collection tubes(1), to stabilize the protein content of blood, enabling better protein biomarker discovery and proteomics experimentation from human blood. The BD P100 tubes contain 15.8 ml of spray-dried K2EDTA and a lyophilized proprietary broad spectrum cocktail of protease inhibitors to prevent coagulation and stabilize the plasma proteins. They also include a mechanical separator, which provides a physical barrier between plasma and cell pellets after centrifugation. Few methods have been devised to extract DNA from clotted blood samples collected in old plasma tubes(2-4). Challenges from these methods were mainly associated with the type of separator inside the tubes (gel separator) and included difficulty in recovering the clotted blood, the inconvenience of fragmenting or dispersing the clot, and obstruction of the clot extraction by the separation gel. We present the first method that extracts and purifies genomic DNA from blood drawn in the new BD P100 tubes. We compare the quality of the DNA sample from P100 tubes to that from EDTA tubes. Our approach is simple and efficient. It involves four major steps as follows: 1) the use of a plasma BD P100 (BD Diagnostics, Sparks, MD, USA) tube with mechanical separator for blood collection, 2) the removal of the mechanical separator using a combination of sucrose and a sterile paperclip metallic hook, 3) the separation of the buffy coat layer containing the white cells and 4) the isolation of the genomic DNA from the buffy coat using a regular commercial DNA extraction kit or a similar standard protocol.

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AccuPrep Genomic DNA Extr Kits for 96 well Plate Va AccuPrep Genomic DNA Extr AccuPrep Genomic DNA Extr AccuzolTM Total RNA Extra Human Dnak (HSP70) His ta ProPrep™ Genomic XL-10 Bone Morphogenetic Protei DNA (cytosine 5) methyltr TOM1-like protein 2 antib Recombinant E. coli HSP70 Recombinant E. coli HSP70

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#23688414   2013/09/02 Save this To Up

Thymic stromal lymphopoietin activity is increased in nasal polyps of patients with chronic rhinosinusitis.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with TH2-dominant inflammation. Thymic stromal lymphopoietin (TSLP) is a cytokine that triggers dendritic cell-mediated TH2 inflammatory responses and that enhances IL-1-dependent TH2 cytokine production in mast cells. Although increased TSLP mRNA levels have been found in nasal polyps (NPs), expression of TSLP protein and its function in patients with chronic rhinosinusitis (CRS) have not been fully explored.

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Human Thymic Stromal Lymp Mouse Anti-Lipoprotein Li Rat monoclonal anti mouse Interleukin-34 IL34 (N-t Interleukin-34 IL34 anti Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon HIV1 integrase antibody, Goat Anti-Human Laforin ( Akt Inhibitor, Isozyme Se

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#8887717   1997/02/11 Save this To Up

Validation and application of an immunoradiometric assay for the determination of human parathyroid hormone fragment 1-34 in dog plasma following subcutaneous and intravenous administration.

A method for the measurement of human parathyroid hormone fragment 1-34 (PTH1-34) in dog plasma was developed by modification of a commercially available immunodiometric assay (IRMA) designed for the determination of rat PTH1-34 in serum. Major modifications were made to the assay in order to circumvent significant problems encountered during the validation of the IRMA. PTH1-34 was found to be highly unstable in both rat serum and dog serum and plasma at room temperature, in contrast to literature reports. The addition of a protease inhibitor cocktail to serum or plasma samples was necessary to prevent in-vitro proteolytic degradation of human PTH1-34 prior to analysis. Additionally, plasma was chosen over serum as the sample matrix to expedite the separation of samples from cells, minimizing proteolytic degradation prior to the addition of cocktail. Finally, the reported 100% cross-reactivity between rat and human PTH1-34 was found to be only 65%; therefore, a human PTH1-34 standard was substituted for the rat standard. These modifications allowed the accurate measurement of human PTH1-34 in plasma obtained from dogs dosed intravenously and subcutaneously with human PTH1-34 using a commercially available kit.

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Rabbit Anti-intestinal FA Mouse Anti-Parathyroid Ho Rabbit Anti-Human Androge Rabbit Anti-Human Androge Integrin β1 (CD29) Antib LPAM-1(Integrin α4, CD49 α-Internexin Antibody So INPP5F antibody Source Ra Interferon alpha-8 antibo Interferon alpha-6 antibo succinate-CoA ligase, GDP interleukin 17 receptor C

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