the structural evolution of side chain grafted polypyrrole to carbon membrane
Macro to micro porous ceramic and carbon media philosophy of design and fabrication
... °C to 1000 °C, with the optimal temperature setting at 800 °C In Chapter 11, The structural evolution of side- chain grafted polypyrrole to carbon membrane , an insight into the evolution of carbon ... consisting of BPO initiator in chloroform was dripped on to the surface of the carbon membrane, allowing the infiltration of this solution into the nano-sized pore channels With the evaporation of chloroform, ... : The Structural Evolution of side- chain grafted Polypyrrole to Carbon Membrane 248 11.1 Introduction 249 11.2 Experimental 252 11.2.1 Synthesis of the...
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Safe carrying methods
... death of an employee If an employer hires a contractor to carry out work then the contractor and any of their employees are deemed to be employees of the employer If an employer fails to protect the ... that they have it skin cancers are the most common cancers in Australia Australia has the highest rate of skin cancers in the world? The longer skin is exposed to the sun, the greater the risk of ... employees they will be in breach of the law and liable for a fine of up to $100 000 If an employer fails to protect the employees and as a result causes the death or serious harm to the employee they...
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Glycoprotein methods protocols - biotechnology 048-9-001-013.pdf
... weight of the sample is calculated from the volume of the tube and the final density of the solution Add sodium phosphate buffer to give the final weight and stir the sample gently Measure the density ... three-fourths of the total volume held by the tube) If required, add 1% CHAPS solution to give a final concentration of 0.01% (i.e., 1% of the total volume) The concentration of guanidinium chloride in the ... and the volume of the CsCl and CHAPS must therefore be compensated for by the addition of a small volume of M guanidinium chloride The final weight of the sample is calculated from the volume of...
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Glycoprotein methods protocols - biotechnology 048-9-027-044.pdf
... to the number of reactive glycol structures Several modifications of the PAS stain have been described These relate to the variable structure of sialic acid and specifically to the presence of ... fixed in solutions other than formalin, e.g., methacarn (methanol:chloroform:acetone, 60:30:10) (44) It is beyond the scope of the current work to discuss the full range of histochemical techniques ... alone to determine whether there is biotin or biotin-like activity in the tissues that should be suppressed prior to the commencement of the immunohistochemistry routine Deglycosylation of mucin...
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Glycoprotein methods protocols - biotechnology 048-9-045-055.pdf
... and the number of samples to be processed Owing to the high O-linked carbohydrate content of mucins (as much as 90% of the total weight), many assays are targeted toward this portion of the molecule ... Protocols: The Mucins (Corfield, T., ed.), Humana, Totowa, NJ Carraway, K L (2000) Preparation of membrane mucin, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T ed.), Humana, Totowa, ... devices) and then the entire membrane in three changes of PBS or TBS Either proceed directly to PAS or detection using antibodies or lectins (see Subheadings 3.2.2 and 3.2.3.) or store the membrane...
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Glycoprotein methods protocols - biotechnology 048-9-057-064.pdf
... one-third of the mucous layer in the stomach and nine-tenths of that in the colon to leave a firm adherent gel, whereas in the duodenum all the mucus can be sucked off It is not yet clear whether the ... luminal surface of the mucus is enhanced visually by the addition of carbon particles to the bathing solution The thickness of the mucous layer is best measured by recording the distance travelled ... gel surface, the extent of which depends on the hydration of exposed mucosa and the degree of stirring of the solution above the mucosa, respectively Consequently, the dimensions of mucous thickness...
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Glycoprotein methods protocols - biotechnology 048-9-065-073.pdf
... the protein concentration of the homogenates: a, e, and i The specific mRNA concentration is calculated relative to the total amount of RNA per cell sample, p The total protein synthesis in the ... significance, largely because of the use of arbitrary units for each of the measurements of radioactivity, owing to the use of the PhosphorImager The identification of mucin precursors and mature ... using the PhosphorImager (see Note 4) Calculate the amount of mature mucin as au/milliliter of homogenate or as au/milliliter of medium Quantify the total amount of the mucin of interest in the...
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Glycoprotein methods protocols - biotechnology 048-9-075-085.pdf
... constitutes 20% of the total mass of the mucin subunit and that approx 50% of this protein is in “naked” regions that are accessible to the proteinase Chromatography separates the high Mr mucin ... variety of lectins and mucin- or carbohydrate-specific antibodies (for a discussion of the relative merits and drawbacks of these analytical tools, see Chapter 4) The homogeneity of the fractions ... dilution) the mucins into reduction buffer Reduce the mucins by the addition of DTT to a final concentration of 10 mM for h at 37°C Alkylate the free-thiol groups generated by reduction with the addition...
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Glycoprotein methods protocols - biotechnology 048-9-087-096.pdf
... 10) Touch a drop (1 µL) of the spreading agent to the surface and leave the solution for 5–15 In this time the mucins diffuse to the surface and become entrapped in the BAC monolayer Touch the carbon ... floating carbon film This allows the intact film to be steered on the water surface over the grids Lower the water level by gentle suction to allow the carbon film to be deposited on the grids Dry the ... that the physical size and shape of the molecule are unchanged by the supporting medium and that the rotor speed is constant during the experiment, then the change in the sedimentation rate of the...
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Glycoprotein methods protocols - biotechnology 048-9-111-119.pdf
... aliquot of a solution of a sample of mucin into an autosampler glass vial and dry under vacuum (see Note 3) Place the vials, 400 µL of HCl (6 N), and a crystal of phenol into the bottom of the hydrolysis ... protocol for PVDF-bound protein samples During the evacuation and argon flush of the hydrolysis vessel, the bottom of the vessel will become cold and the acid will boil under vacuum These are the ... where is the dilution factor in derivatization bThe pmol of mucin amino acid = peak area of mucin amino acid/peak area of standard amino acid × 25 pmol × 2, where is the dilution factor from the half...
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Glycoprotein methods protocols - biotechnology 048-9-121-128.pdf
... support, the sialic acid must be removed to prevent the formation of an amide bond between the carboxyl group of the sialic acid and the amine of the support Although the alternative of using ... proteins because of the coupling of the χ- and δ-carboxyl groups to the membrane Acknowledgments NEZ was supported by an Australian Postgraduate Award AAG acknowledges the support of the National ... Dry the membrane after all of the sample has been applied Mix approx mg of EDC in 50 µL of reagent attachment buffer (see Note 15) and carefully pipet 10–50 µL onto the arylamine-derivatized membrane...
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Glycoprotein methods protocols - biotechnology 048-9-129-141.pdf
... assembled on to the block according to the synthesis schedule It is important that only gears requiring deprotection in the next cycle of synthesis be added to the block Achieve deprotection of the amino ... 13) The addition of bromophenol blue to the HOBt to give a final concentration of 0.05 mM as an indicator of coupling efficiency is optional The volume of HOBt solution specified on the synthesis ... volumes of the various reagents required on each day of the synthesis and then instruct the operator where in the 96-well reaction tray each amino acid should go (see Notes 5–8) 3.2.2 Peptide Synthesis...
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Glycoprotein methods protocols - biotechnology 048-9-143-155.pdf
... necessary to establish the ideal ratio of DNA to Lipofectamine For this system, add µg of pRMC or pSVL control to 100 µL of DMEM Add 10 µL of Lipofectamine to 100 µL of DMEM in a separate tube Add the ... dishes of COS cells and wash with mL of DMEM (containing no antibiotics or FBS) Drain the dishes thoroughly Add 800 µL of DMEM to the DNA-Lipofectamine mixture, pipet to mix, and add to the dish of ... Wash the COS cells in PBS, scrape the cells into 500 µL of PBS and microfuge the suspension at 4˚C to form a pellet Resuspend the pellet in 50 µL of 1% SDS, and boil to denature Dilute the suspension...
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Glycoprotein methods protocols - biotechnology 048-9-159-180.pdf
... these conditions, the hydroxyl groups of carbohydrates are converted to oxyanions with pKa values in the range of 12–14 The anomeric hydroxyl group of the reducing sugar is more acidic than the ... (complete dehydration of samples is the main condition of success) amounts of glycoproteins or oligosaccharides corresponding to 10 µg of total sugar to which µg of mesoinositol is added as an internal ... Trimethylsilylate with 20 µL of BSTFA in the presence of 10 µL of pyridine for h at room temperature Apply 1–5 µL of the solution of trimethylsilylated methylglycosides to GLC 3.2.4.2 GAS CHROMATOGRAPHY CONDITIONS...
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Glycoprotein methods protocols - biotechnology 048-9-181-190.pdf
... retention times to each other and to their alditols On HPAEC (13), the reducing oligosaccharides will be retained significantly longer than the alditols, to give improved chromatography Sulfated ... microscale identification of the presence of alditols, dissolve the sample in 200 µL 0.2 M NH4OH and add to the top of a PBA minicolumn prewashed with MeOH, water, and 0.2 M NH4OH Wash the PBA column with ... 0.2 M of NH4OH and × 100 µL of water Specifically elute the alditols in M acetic acid Evaporate the sample and reevaporate with × 100 µL of water 3.2 Oxidation of Oligosaccharide Alditols To the...
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Glycoprotein methods protocols - biotechnology 048-9-191-209.pdf
... consists of the identification of the core region (Table 1), based on the characteristic chemical shifts of the H-2 and H-5 atom resonances of the GalNAc-ol unit Moreover, the quadruplet of the H-6' ... establishing the sequence of carbohydrate chains Since the strongest NOE is not always between the protons connected to the linkage, the method may fail to establish the position of the glycosidic ... N-acetylgalactosaminitol results in two fragments: one corresponds to the 3-branch and the other to the 6-branch of the Analysis of Mucin-Type O-Linked Oligosaccharides 207 Fig ESI-MS of the 3-branch,...
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Glycoprotein methods protocols - biotechnology 048-9-211-216.pdf
... for the analysis of inorganic ions The limit of detection of sulfate by ion chromatography as described here is about nmol Since mucins are often highly sulfated (i.e., the number of moles of ... elution positions of ionic species using this method Attach the AS11 ion column to the high-performance liquid chromatograph Attach the ion suppressor downstream of the column, and the conductivity ... downstream of the ion suppressor, according to the manufacturer’s recommendations Attach the neutralizing counterflow (50 mM sulfuric acid) reservoir to the ion suppressor The initial chromatographic...
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Glycoprotein methods protocols - biotechnology 048-9-219-226.pdf
... upper half of the crypts, where they differentiate These differentiated cells migrate to the top of the crypt, and there is evidence that they die by apoptosis and that apoptosis may be the terminal ... into the medium, although some of the mucin may remain adherent to the epithelial tissue The submerged culture system, under basic nonstimulated conditions, leads to the recovery of 20% of the ... guish them from the cells in vivo On the other hand, the cell lines may consist of only one cell type, which can be an advantage Study of the colon has the advantage that the sequence of changes...
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Glycoprotein methods protocols - biotechnology 048-9-227-237.pdf
... However, the possibilities of metabolic labeling to study minute amounts of mucin can also be applied to the mucin precursors, i.e., to the biosynthesis of the mucin polypeptide and other early ... gradient Aspirate 0.5-mL samples from the top of the tube by pipet, or drain from the bottom after piercing the tubes Weigh the samples to obtain the density of each fraction Slot-blot aliquots ... migration of the sample into the gels (8) 17 Markers for agarose gel electrophoresis reflect the separation of molecules on the basis of their size and charge This is in contrast to the normal...
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