The effect of sonication temperature on the debundling of carbon nanotube (CNT) macro-bundles is reported and demonstrated by analysis with different particle sizing methods. The change of bundle size over time and after several comparatively gentle sonication cycles of suspensions at various temperatures is reported. A novel technique is presented that produces a more homogeneous nanotube dispersion by lowering the temperature during sonication. We produce evidence that temperature influences the suspension stability, and that low temperatures are preferable to obtain better dispersion without increasing damage to the CNT walls.
Hi, just a general query: how do other people sonicate samples after protein overexpression? In one lab a postdoc divided samples into 5ml or bigger pots and sonicated on maximum for for 20 seconds with a minutes between each sonication 2-3 times (on ice). In my present lab people put their whole sample in one 50ml pot and sonicate for longer e.g. 30-60 seconds and more times e.g. 4-5. Is one way better than the other, is it even important as long as the cells change consistancy and lyse completely and do not warm up? I have started sonicating in 1 big pot for 20 seconds with 1-2 minute intervals as in the method above I think the sample starts to heat up. No-one I have asked seems that sure about this, I was wondering if anyone on the forum does...?. ...
1. Tissue: Cut 25mg of tissue into small pieces, place in 1.5ml centrifuge tube and and 180 l buffer T1.. Liquid nitrogen samples: Transfer the powder from 1cm of tissue to a 1.5ml centrifuge tube and add 180 l buffer T1.. Mechanical homogenized samples (Polytron, Ultra Turrax): Add 25mg of tissue to a 1.5ml centrifuge tube and add 180 l PBS buffer. After homogenization proceed with step 3.. 2. Make sure that buffers B3, B5 and the proteinase K solution have been prepared according to the remarks above.. 3. Add 25 l of proteinase K stock solution, mix by vortexing, and incubate at 56C in a shaking waterbath until complete lysis is obtained (~1-3h). Additional 3-4 times vortexing every 10-15 minutes leads to shorter lysis times. At the end of this incubation prepare a 70C waterbath.. 4. Add 200 l buffer B3 to the sample, vortex the mixture and incubate it at 70C for 10 minutes.. 5. Add 210 l ethanol to the sample and vortex immediately. If a white precipitate is obtained after the addition of ...
I am a beginner in western blot. is it possible to vortex antibodies (primery &secondary) upon dilution in 3% skim milk ? or is it better to mix by pipetting up and down?. ...
In article ,32F88C97.1431 at sheffield.ac.uk,, K.Mulcahy at sheffield.ac.uk wrote: However, one of my plasmids is 10.5 , kb. Does anyone know if plasmids (pre-linearised) which are this large , can be shaken vigorously without causing cleavage of the linear DNA , fragment? If not, then what is the best way to extract them with , phenol:chloroform? , There is no upper limit as long as there is no standard for vigorous shaking by hand. If you are worried about breakage, try Centriflex-AG cartridge from AGTC: all you need is to centrifuge your sample through it , protein is trapped in the cartridge. Tel 001-301-990-2685, Cat No 10319. No affiliation. -- Alexander Kraev, PhD Biochemie III, ETHZ Zurich Phone 41-1-632-31-47 Fax 41-1-632-12-13 e-mail kraev at bc.biol.ethz.ch ...
If you follow the sonication protocol below for cell lysis you should achieve efficient lysis of your cells for your required application. Tips Included.
We compared the properties of the nuclei that accumulate in 7.5 mM-KCl in ATP-G-actin solutions and of the oligomers that are formed by sonication of either G-actin or F-actin. We found that the ability of the above species to prime the polymerization of actin decays with different rates. The nuclei are stable in 7.5 mM-KCl (they decay with a rate constant of 1.5 X 10(-3) s -1 at pH 7.8 at 22 degrees C in the absence of KCl). The oligomers formed by sonication of either G-actin or F-actin, once the sonication is stopped, revert to simpler structures or evolve into F-actin, depending on the KCl concentration in which they are kept. In 10.5 mM-KCl at pH 7.8 at 22 degrees C their priming ability decays with a rate constant of 6 X 10(-3) s -1. We propose that the nuclei that form spontaneously in 7.5 mM-KCl are not directly susceptible to elongation. They must first be converted into activated nuclei, which exist in very low concentration at the steady state. The activated nuclei are directly ...
In article ,2lo72q$9ur at news.iastate.edu,, bipin at iastate.edu (Bipin K Dalmia) writes: ,, ive never had much success with sonication but since that is the only ,, piece of equipment we have at present, do you have any suggestions for ,, optimizing e. coli disruption with a sonicator? ive heard that some ,, people use lysozyme treatment before sonicating, do you have any ,, experience with this?? any other treatments?? ,, ,, bip ,, -- ,, bipin k. dalmia the other night i was lying on my bed, looking ,, bipin at iastate.edu up at the beautiful stars, and i said to myself, ,, n2.bkd at isumvs.iastate.edu where the F*CK is my ROOF !! ,, -- Ive used sonication, with and without lysozyme, and also freeze-thawing to lyse coli (freeze-thawing as in multiple cycles of flash freezing in liquid N2, then rapid thawing in a warm water bath (we use 65C, but dot let the samples get to 65, or much above 30 really, and only invert gently) Ive had a _lot_ more sucess with sonication than with freeze- ...
Foar it ûntstean fan astaxanthin (AX), de kombinearre behanneling fan TPP en sonication mei Hielschers UP400S is net allinnich ienfâldige ans maklik te brûken, mar ek tige effisjinte technyk om AX te izeren fan de baktearjende biomass fan Paracoccus NBRC 101723. De operative fariabelen fan sonication kinne optimisearre wurde om optimale ekstra-resultaten te krijen. Uterrasjonêre ferwurkingsparameters binne ek as amplitude, tiid, temperatuer en ekstra skip (groei, foarm), lykas de partikelgrutte fan biomass, ynfloed op de ultrasonike frijlitting fan AX fan biomass. Ultrasonication fan wiete biomasse jout bêste resultaten as de ultrasjonale TPP fan droege biomass foar flugge en effisjoneare ekstraksje fan AX. De ultrasone TPP resultaat yn in 37% heger AX-herstel as de konvinsjonele ekstrawinning. (Chougle et al., 2013). ...
Description: When required for use as a vaccine antigen, H. pylori was harvested from plates with 0.1 M phosphate-buffered saline (PBS), pelleted by centrifugation, and disrupted by sonication. The sonicate was stored at 220°C until use. Purified catalase was obtained by the method of Hazell et al. (11). Briefly, H. pylori cells were harvested with 0.1 M sodium phosphate buffer (pH 7.2), centrifuged, and then resuspended in the buffer. Cells were disrupted by sonication, cellular material was removed by centrifugation (5 min, 10,000 3 g), and then the supernatant was collected and filtered (0.22-mm-pore-size filter). Catalase was eluted by the creation of a gradient with 1 M NaCl in 0.01 M sodium phosphate buffer. The purified catalase was then filter sterilized, stored at 4°C, and protected from light (Radcliff et al., 1997 ...
Differential spectra of the reduced minus oxidized ingredients were recorded on a beam/double wavelength spectrophotometer. The maxima intake for cyt b and for cyt c c1 used were 561 and 550 nm, respectively. The cyt c/cyt b ratio was always used to change the full total protein content in the different examples. As defined in ref. immunoprecipitation was performed utilizing the IP50 kit from Sigma. Shortly, cells were ressuspended in buffer supplemented Capecitabine solubility using a blend of protease and phosphatase inhibitors. Cells were broken routinely by vortexing with glass beads, and 100 ul of 10? lysis buffer was added to 1 ml of cell suspension and incubated at 4 C during 1 h. 2 ug of monoclonal anti Bax antibody was added, and the lysate incubated over night at 4 C. Protein G paired agarose beads were added and incubated for 6 h. Washing and recovery of the samples were done following manufacturers instructions. Similar samples were packed in parallel onto two SDS PAGE ties in and ...
Cell culture, transfection, immunocytochemistry, and Western blot. Calcium phosphate transfections, immunocytochemistry, and culture of dissociated hippocampal CA3/CA1 pyramidal neurons were performed as described previously (Wu et al., 2001). Enhanced green fluorescent protein (EGFP) was cotransfected with hemagglutinin (HA)- or myc-tagged constructs to visualize the detailed cell morphology. Hippocampal slice culture was done from 7-8 d postnatal rats and transfected biolistically at 2 d in vitro (DIV) as described previously (Lo et al., 1994). A modified procedure was used for Western blot experiments using cells growing on 12 mm coverslips. In brief, coverslips were placed in 1.5 ml microcentrifuge tubes containing 40 μl of 9 m urea and were crushed, and, after vigorous vortexing, samples were centrifuged (18,000 × g for 2 min). The protein concentration was determined by BCA assay (Pierce, Rockford, IL), and volumes were adjusted with 9 m urea so that all samples were 0.3-1.0 μg/μl. ...
We recommend that you treat the amplified DNA exactly as you would treat genomic DNA. The best way to store gDNA is frozen at high concentration at -20ºC in 1x TE. The worst way to store gDNA or the amplification product is at low concentration (less than 10 ng/ul) at 4ºC in water.. We have observed significant degradation of genomic DNA stored at 2-3ng/ul at 4ºC in as little as 2-3 weeks. Frozen samples can be thawed and aliquoted. Keep a working stock to use directly in any downstream application. This will decrease the risk for degradation of higher concentrated stocks.. Repeated freeze-thaws, vortexing, rapid pipetting up and down, storage in water and heating to over 75ºC should be avoided as these processes can shear gDNA or MDA amplified DNA, and/or contribute to rapid DNA degradation.. ...
Large Ampule/Tube Attachment [ESI-0511] - The Large Ampule/Tube Attachment can be used with the Vortex-Genie 2 family of mixers and the Vortex-Genie Pulse. It provides vigorous end-to-end agitation combined with vortexing of up to four 10mm-17mm diameter tubes or ampules.
ExiSpin™ has the functions of a vortexer as well as a microcentrifuge and will significantly save your time in the lab. ExiSpin™ comes complete with rotor for 4 x 8-strip tubes and 12 x 1.5/2.0 ml tubes. Use it for Minipreps, to resuspend oligos, set up your PCR reaction, or any task that requires vortexing in micro test tube or 0.2 ml PCR tubes ...
In this method order clomid australia womens health clinic elizabeth, dried lipids are homogenized with an aqueous solution containing the enzyme to be encapsulated order clomid with paypal breast cancer 5k topeka ks, frozen buy clomid pregnancy 38 weeks, and lyophilized discount cialis 2.5mg line. The lyophilized powder is then hydrated in one-tenth of the starting volume of the liposome dispersion discount viagra 75mg without a prescription, gen- tly stirred, and completed with the rest of the volume after a hydration step (21,32). The fate of liposomes in vivo after intravenous administration is dependent on several factors, namely, lipid composition, surface charge, steric effect, fluidity of the lipid bilayer, and mean size of liposomes. Sonication is a mechanical method in which liposomal suspension is subjected to ultrasound by using either sonication probe or sonica- tor bath. This method is not appropriate for the encapsulation of enzymes in liposomes, as it results in low loadings ...
Those looking for under-the-radar talent should check out Damien the Demon for a set of eclectic beats. From the sparse almost monochromatic drum-driven beat of "Long Daze" to the screaming distorted electronics of "Fire Ball", Damien the Demon for moves sonically from A to Z without blinking a stylistic eyelid. Production is undoubtedly one of biggest aspects of any Hip-Hop song and it has the ability to make or break a record. Since we are so technologically advanced in 2015, producers have the ability to produce almost anything imaginable.. And thats exactly what Damien the Demon does. With no boundaries or limits, he throws together sounds more than styles. You may, or may not like what comes out of his musical blender, but its original and off the beaten track. Id go as far to say that Damiens beats are an acquired taste and definitely not for everybody. Hiphop as a genre is in a perpetual state of forward motion. Just as there will always be people yearning for the good old days, there ...
JoVE publishes peer-reviewed scientific video protocols to accelerate biological, medical, chemical and physical research. Watch our scientific video articles.
YidC, a known person in the YidC/Oxa1/Alb3 family members, inserts proteins in to the membrane and facilitates membrane-protein folding in bacterias. 19 and 14 residues accompanied by the GFP-His8 label had been taken off YidC to create YidC27C266 and YidC27C261, respectively (Fig. 1 ? (TEV) protease … 2.2. Fluorescent size-exclusion chromatography (FSEC) ? FSEC was performed as defined previously with adjustments (Kawate & Gouaux, 2006 ?). The C-terminally GFP-His8-tagged YidC proteins had been overproduced in C41(DE3) or BL21(DE3) cells harbouring pRARE (Novagen) as well as the pCGFP-BC-based plasmid under a number of growth circumstances by changing essential parameters such as for example lifestyle temperature, induction and duration timing. The cells had been harvested in 5?ml LB moderate supplemented with appropriate antibiotics. The cells had been harvested, resuspended in buffer (20?mTrisCHCl pH 8.0, 300?mNaCl, 0.1?mphenylmethylsulfonyl fluoride) and disrupted by sonication using a ...
YidC, a known person in the YidC/Oxa1/Alb3 family members, inserts proteins in to the membrane and facilitates membrane-protein folding in bacterias. 19 and 14 residues accompanied by the GFP-His8 label had been taken off YidC to create YidC27C266 and YidC27C261, respectively (Fig. 1 ? (TEV) protease … 2.2. Fluorescent size-exclusion chromatography (FSEC) ? FSEC was performed as defined previously with adjustments (Kawate & Gouaux, 2006 ?). The C-terminally GFP-His8-tagged YidC proteins had been overproduced in C41(DE3) or BL21(DE3) cells harbouring pRARE (Novagen) as well as the pCGFP-BC-based plasmid under a number of growth circumstances by changing essential parameters such as for example lifestyle temperature, induction and duration timing. The cells had been harvested in 5?ml LB moderate supplemented with appropriate antibiotics. The cells had been harvested, resuspended in buffer (20?mTrisCHCl pH 8.0, 300?mNaCl, 0.1?mphenylmethylsulfonyl fluoride) and disrupted by sonication using a ...
Wash off unbound antibody: pellet beads (3000 rpm/ 4°C/ 3 min.), discard sup, wash w/ complete sonication buffer (keep on ice); repeat ...
MethylCollector Ultra is a fast magnetic assay to enrich for CpG-methylated DNA from cell or tissue samples fragmented by sonication or enzymatic digestion; these assays provide more specific enrichment of methylated DNA than antibody-
Do not have have gel image for 2nd shearing attempt (4 additional cycles), but looked better than first. Because final attempt with additional 2 cycles did not look right (more large fragments), decided to continue with subsequent steps to avoid further sonication problems. ...
Howdy folks. This is my attempt to spotlight some lesser known tidbits of psychedelia. The aim is to sonically reprogram your mind. Im always searching for something new and bizarre, so check back every once and a while. And if you have any suggestions or mp3s of something really interesting, please drop me a line ...
Our Microfluidizer cell disruption equipment allow for effective, efficient cell lysis in laboratory and production applications. Learn more -- request details.
TY - JOUR. T1 - Larynx Decellularization. T2 - Combining Freeze-Drying and Sonication as an Effective Method. AU - Hung, Shih Han. AU - Su, Chin Hui. AU - Lee, Fei Peng. AU - Tseng, How. PY - 2013/5. Y1 - 2013/5. N2 - Objectives: Ideal methods for the reconstruction of the laryngeal structure and restoration of the laryngeal function once the larynx has been damaged or removed have not yet been developed. Thus, larynx tissue engineering practices have recently been extensively investigated. A scaffold may be generated using biocompatible or artificial materials. Decellularization methods, which use preexisting tissues as material sources, have also been used to manufacture larynx scaffolds with promising results. In this study, we developed a novel decellularization method that combines freezing, drying, and sonication. Study Design: Porcine model study. Methods: Fresh porcine larynxes were used for decellularization. The process of the decellularization cycle comprised overnight freeze-drying, ...
The objective of this Phase IV study is to evaluate the safety of the ExAblate treatment of uterine fibroids using the enhanced sonication techniques, based on the current commercially-approved treatment guidelines. Treatment may include up to 100% of individual fibroid volume, within established serosal and sacral treatment margins.. The Enhanced sonication is one of the various sonication modes that may lead to increased thermal dose volume of each sonication without additional safety risks. This is an additional treatment tool available in the ExAblate system for the treatment of uterine fibroids.. The safety profile of the Enhanced Sonication was investigated under an FDA-regulated IDE study. FDA granted approval of Enhanced Sonication with the requirement to perform a post-approval study to collect additional safety data when treating up to 100% of individual fibroid volume. ...
To continue my blog part 1 (Part 1:https://blog.restek.com/?p=67087) , where I have briefly discussed the importance of separating the d and l isomers to accurately identify the illicit isomer using an achiral method on a Raptor C18 column employing a pre-column derivatization technique. Today Id like to discuss more about the matrix of interest, sample preparation and derivatization, chromatographic and mass spectrometric conditions.. Sample Preparation: 50 ?L of calibration standard or QC sample prepared in analyte free pooled human urine (spiked in the range of 50-5000ng/mL) was aliquoted into a micro centrifuge tube. 10 ?L of a working internal standard (20 ?g/mL (±)-amphetamine-D11 and (±)-methamphetamine-D11 in water) and 20 ?L of 1M NaHCO3 was added and vortexed at 3000 rpm for 10 seconds. After vortexing, 100 ?L of 0.1% (w/v) Marfeys reagent (1-fluoro-2-4-dinitrophenyl-5-L-alanine amide) prepared in acetone was added, vortexed, and heated at 45 °C for 1 hour in a water bath. Samples ...
After removal in the operating room, implants are placed in the air-tight container and transported to the microbiological laboratory. After addition of Ringers solution, the implant is processed by vortexing (30 seconds) and sonication (1 minute) to dislodge (planktonize) microorganism into the surrounding fluid (sonicate). The sonication fluid is cultured on aerobic and anaerobic agar plates and inoculated in broth media.. ...
The purpose of this investigation was to quantify the effects of storage temperature, duration, and the urinary sediment on urinary hydration markers. Thirty-six human urine samples were analyzed fresh and then the remaining sample was separated into 24 separate vials, six in each of the following four temperatures: 22 °C, 7 °C, -20 °C, and -80 °C. Two of each sample stored in any given temperature, were analyzed after 1, 2, and 7 days either following vortexing or centrifugation. Each urine sample was analyzed for osmolality (UOsm), urine specific gravity (USG), and urine color (UC). UOsm was stable at 22 °C, for 1 day (+5-9 mmol∙kg-1, p , .05) and at 7 °C, UOsm up to 7 days (+8-8 mmol∙kg-1, p , .05). At -20 and -80 °C, UOsm decreased after 1, 2, and 7 days (9-61 mmol∙kg-1, p , .05). Vortexing the sample before analysis further decreased only UOsm in the -20 °C and -80 °C storage. USG remained stable up to 7 days when samples were stored in 22 °C or 7 °C (p , .05) but declined ...
We examined MCE to evaluate myocardial perfusion. MCE is somewhat limited when used to evaluate myocardial perfusion quantitatively, although it accurately demonstrates the area at risk ([22-25]). Quantitative evaluation of myocardial perfusion using the contrast echo technique requires use of a contrast agent that distributes bubbles of the same density and size throughout all injections and among different subjects, because the echo intensity significantly depends on bubble size and density. There are several studies presenting indexes from the time-intensity curve that are reproducible and reliable when human albumin is used as the contrast agent, bubbles are produced by a sonication method, the contrast solution has no foamy layer and the injection volume and speed are carefully set ([26-36]). Recent work ([37, 38]) has suggested that the ultrasonic power and the gas surrounding the bubbles are major factors in the disruption of bubbles, although these factors have not yet been well ...
The purpose of the present research is the different morphologies production of crystalline and amorphous-silica powder. Its a basic material for many pharmaceutical and environmental applications as well. And, its produced using the combination of the alkali chemical etching process and the ultra-sonication technique. The critical preparation conditions are KOH concentration (weight %) and the sonication time (hour). The paper presents the chemical mechanism of the silica particle formation as well as the different morphology. The results show the formation of crystalline and amorphous-porous-silica particles in the micrometer range with the porous order network that has pore sizes range in micrometer too. This synthetic uses commercial silicon, which could be useful for large-scale production. Also, the nano-sphere and nano-cubic shapes of silica powder are formed starting by commercial silicon powder.
0159]a) Silanization of the nanoparticles by the use of a hetero bifunctional silane with a subsequent further functionalization, e.g. PEG-ylation (two-step PEG-ylation procedure). Filtered nanoparticles are sonicated for 15 minutes, in order to break agglomerates. 1 ml of the nanoparticles in a water suspension (typically dialysed in a previous step) is then placed in an eppendorf tube and 50 μl of the bifunctional silane, e.g. 3-aminopropyl triethoxy silane, is added followed by vortexing and 1 h of sonication. During the reaction the silane function binds to the surface of the nanoparticles leaving the other function, e.g. an amino function, free for the subsequent functionalization step, e.g. introduction of hydrophilic polymers such as polyethylene glycol (PEG-ylation). If needed the silane is added together with a solvent with due care taken for favouring reaction between silane and nanoparticles compared to polymerisation of the silane. 10 μL of Milli-Q is then added whereafter the ...
We demonstrate the ability of a staphylococcal nuclease modified with the DsbA periplasm export signal (BBa_K729004) to prevent transfer of genetically modified material to a commercially competent TOP10 E. coli strain. This observation is consistent with our proposed function of BBa_K729019 to act as a biosafety mechanism preventing uptake of released DNA by naturally competent bacteria. We used sonication to disrupt three cell types: unmodified W3110 strain E. coli and W3110 strains harbouring chloramphenical resistance plasmids encoding a laccase (BBa_K729006) or the DsbA-Nuclease (BBa_K729019). All three strains were grown to OD600= 2.0 prior to sonication, in 2mL LB broth which contained 100ug/mL chloramphenical for plasmid-harbouring cells. After sonication, disruptates were incubated for 10min at room temperature. 5uL of the disruptate was used to transform an aliquot of TOP10 chemically competent cells, following manufacturer instructions. As a control, we also spread 20uL of the ...
We demonstrate the ability of a staphylococcal nuclease modified with the DsbA periplasm export signal (BBa_K729004) to prevent transfer of genetically modified material to a commercially competent TOP10 E. coli strain. This observation is consistent with our proposed function of BBa_K729004 to act as a biosafety mechanism preventing uptake of released DNA by naturally competent bacteria. We used sonication to disrupt three cell types: unmodified W3110 strain E. coli and W3110 strains harbouring chloramphenical resistance plasmids encoding a laccase (BBa_XXXXXX) or the DsbA-Nuclease (BBa_K729004). All three strains were grown to OD600= 2.0 prior to sonication, in 2mL LB broth which contained 100ug/mL chloramphenical for plasmid-harbouring cells. After sonication, disruptates were incubated for 10min at room temperature. 5uL of the disruptate was used to transform an aliquot of TOP10 chemically competent cells, following manufacturer instructions. As a control, we also spread 20uL of the ...
To continue my blog part 1 (Part 1:https://blog.restek.com/?p=67087) , where I have briefly discussed the importance of separating the d and l isomers to accurately identify the illicit isomer using an achiral method on a Raptor C18 column employing a pre-column derivatization technique. Today Id like to discuss more about the matrix of interest, sample preparation and derivatization, chromatographic and mass spectrometric conditions.. Sample Preparation: 50 ?L of calibration standard or QC sample prepared in analyte free pooled human urine (spiked in the range of 50-5000ng/mL) was aliquoted into a micro centrifuge tube. 10 ?L of a working internal standard (20 ?g/mL (±)-amphetamine-D11 and (±)-methamphetamine-D11 in water) and 20 ?L of 1M NaHCO3 was added and vortexed at 3000 rpm for 10 seconds. After vortexing, 100 ?L of 0.1% (w/v) Marfeys reagent (1-fluoro-2-4-dinitrophenyl-5-L-alanine amide) prepared in acetone was added, vortexed, and heated at 45 °C for 1 hour in a water bath. Samples ...
To continue my blog part 1 (Part 1:https://blog.restek.com/?p=67087) , where I have briefly discussed the importance of separating the d and l isomers to accurately identify the illicit isomer using an achiral method on a Raptor C18 column employing a pre-column derivatization technique. Today Id like to discuss more about the matrix of interest, sample preparation and derivatization, chromatographic and mass spectrometric conditions.. Sample Preparation: 50 ?L of calibration standard or QC sample prepared in analyte free pooled human urine (spiked in the range of 50-5000ng/mL) was aliquoted into a micro centrifuge tube. 10 ?L of a working internal standard (20 ?g/mL (±)-amphetamine-D11 and (±)-methamphetamine-D11 in water) and 20 ?L of 1M NaHCO3 was added and vortexed at 3000 rpm for 10 seconds. After vortexing, 100 ?L of 0.1% (w/v) Marfeys reagent (1-fluoro-2-4-dinitrophenyl-5-L-alanine amide) prepared in acetone was added, vortexed, and heated at 45 °C for 1 hour in a water bath. Samples ...
To continue my blog part 1 (Part 1:https://blog.restek.com/?p=67087) , where I have briefly discussed the importance of separating the d and l isomers to accurately identify the illicit isomer using an achiral method on a Raptor C18 column employing a pre-column derivatization technique. Today Id like to discuss more about the matrix of interest, sample preparation and derivatization, chromatographic and mass spectrometric conditions.. Sample Preparation: 50 ?L of calibration standard or QC sample prepared in analyte free pooled human urine (spiked in the range of 50-5000ng/mL) was aliquoted into a micro centrifuge tube. 10 ?L of a working internal standard (20 ?g/mL (±)-amphetamine-D11 and (±)-methamphetamine-D11 in water) and 20 ?L of 1M NaHCO3 was added and vortexed at 3000 rpm for 10 seconds. After vortexing, 100 ?L of 0.1% (w/v) Marfeys reagent (1-fluoro-2-4-dinitrophenyl-5-L-alanine amide) prepared in acetone was added, vortexed, and heated at 45 °C for 1 hour in a water bath. Samples ...
To continue my blog part 1 (Part 1:https://blog.restek.com/?p=67087) , where I have briefly discussed the importance of separating the d and l isomers to accurately identify the illicit isomer using an achiral method on a Raptor C18 column employing a pre-column derivatization technique. Today Id like to discuss more about the matrix of interest, sample preparation and derivatization, chromatographic and mass spectrometric conditions.. Sample Preparation: 50 ?L of calibration standard or QC sample prepared in analyte free pooled human urine (spiked in the range of 50-5000ng/mL) was aliquoted into a micro centrifuge tube. 10 ?L of a working internal standard (20 ?g/mL (±)-amphetamine-D11 and (±)-methamphetamine-D11 in water) and 20 ?L of 1M NaHCO3 was added and vortexed at 3000 rpm for 10 seconds. After vortexing, 100 ?L of 0.1% (w/v) Marfeys reagent (1-fluoro-2-4-dinitrophenyl-5-L-alanine amide) prepared in acetone was added, vortexed, and heated at 45 °C for 1 hour in a water bath. Samples ...
Expression of DNC. Human DNC with C-terminal His-tag expression was constructed as reported previously (Dolce et al., 2001). DNC protein was expressed in Escherichia coli BL21(DE3) and was purified as described previously (Dolce et al., 2001). The dNTP transport activities of DNC were also assayed as described previously (Dolce et al., 2001).. Reconstitution of DNC. Liposomes were prepared by the size extrusion (MacDonald et al., 1991). Desired amounts of cholesterol or cardiolipin were added to egg yolk or soybean phosphatidylcholine (Avanti Polar Lipids, Alabaster, AL) in chloroform. Mixtures were dried in vacuum overnight. The dried lipid was dissolved in buffer (20 mM PIPES, pH 6.8, 1 mM EDTA) to a final concentration of 10% (w/v) by intensely vortexing. The mixture was frozen (liquid nitrogen) and thawed (37°C water bath) three times and passed through a 100-nm polycarbonate membrane filter 13 times. Proteoliposomes were generated by the detergent removal method (Palmieri et al., 1995). ...
Wittig reaction under Phase Transfer conditions was performed in a flow reaction system. Different bases, aldehydes, phosphonium salts, and flow reaction parameters were investigated, in absence of a phase transfert catalyst. An improvement of the reaction outcome (yield and reaction time) was achieved through the immersion of the reactor into an ultrasound bath. ...
Diagenode has developed advanced sonicators - the Picoruptor® for ChIP-PCR,ChIP-seq and NGS, Megaruptor® for tightest distribution of long DNA fragmentation and the One® which provides small volume DNA shearing for Next-Generation-Sequencing(NGS).
Usono eases, improves and innovates ultrasound use. The ProbeFix improves reproducibility of a continuous ultrasound image for cardiac and intensive care
TY - JOUR. T1 - Ultrasonically assisted fabrication of vaterite submicron-sized carriers. AU - Svenskaya, Yu I.. AU - Fattah, H.. AU - Zakharevich, A. M.. AU - Gorin, D. A.. AU - Sukhorukov, G. B.. AU - Parakhonskiy, B. V.. PY - 2016/3/1. Y1 - 2016/3/1. N2 - Ultrasonically accomplished precipitation of calcium carbonate was demonstrated as a novel beneficial method for the formation of porous submicron vaterite particles with low dispersity, high mass yield, and the "scaling up" possibility. The effect of sonication on the size and crystallographic phase of formed particles was investigated by comparing this method with a magnetic stirring and non-stirring interfusion. By virtue of their porosity, vaterite particles allow the different substance incorporation. The loading properties of synthesized particles were demonstrated for high molecular weight substance (labeled protein), as well as for low molecular one (fluorescent dye). Such features open up perspectives of drug encapsulation and ...
We called this assay: THE LAST RESOURCE =s Basically, we grew cells until an O.D. (600nm) between 0.5-1.0. We needed chubby healthy happy exponential-growth cells because most of our constructions are meant for protein production in this growth phase. Once, we got a lot of cells... we disrupted them by sonication and we analyze the NADH production by their guts using a simple spectrophotometrical analysis. If our parts are working, we should see a higher NADH production when the substrate (dodecanol-1 or dodecanal) to the buffer with microbial guts and NAD buffer. We can use this method because NAD is the natural co-factor of our proteins. Fortunately for us, NAD reduction could be easily quantified by absorbance measurements at 340 nm. We called this protocol "the last resource" because the others didnt work. Which means that alcohols and aldehydes do not cross the cell membrane, thus in order to grow cell on these compounds as sole carbon sources... WE NEEDED TO ADD TRANSPORTERS =( ...
View Notes - W3OutlineLec7 from BIOL D103 at UC Irvine. • Thymosin-Beta4 Sequesters ATP-actin Have plenty of ATP-actin BUTbut it would be
Selection of the suitable SONOREX SUPER depends on the sizes and numbers of objects to be cleaned and of the sample vessels to be sonicated. Different sizes of ultrasonic baths starting from 0.9 to 58.0 litres are available for various applications.Features:PZT-large area oscillating systemsHF-Frequ.... ...
TY - JOUR. T1 - Therapeutic arteriogenesis by ultrasound-mediated VEGF165 plasmid gene delivery to chronically ischemic skeletal muscle. AU - Leong-Poi, Howard. AU - Kuliszewski, Michael A.. AU - Lekas, Michael. AU - Sibbald, Matthew. AU - Teichert-Kuliszewska, Krystyna. AU - Klibanov, Alexander L.. AU - Stewart, Duncan J.. AU - Lindner, Jonathan. PY - 2007/8. Y1 - 2007/8. N2 - Current methods of gene delivery for therapeutic angiogenesis are invasive, requiring either intraarterial or intramuscular administration. A noninvasive method of gene delivery has been developed using ultrasound-mediated destruction of intravenously administered DNA-bearing carrier microbubbles during their microcirculatory transit. Here we show that chronic ischemia could be markedly improved by ultrasound-mediated destruction of microbubbles bearing vascular endothelial growth factor-165 (VEGF165) plasmid DNA. Using a model of severe chronic hindlimb ischemia in rats, we demonstrated that ultrasound mediated ...
160 patients who underwent revision or re-revision total hip or knee arthroplasty for different reasons in our institution were recruited from August 2013 to August 2016. 80 patients meet the criteria of PJI from the muscle and skeletal muscle Association (MSIS) are divided to the infection group, another 80 patients are divided into the non-infection group.. Medical and demographic data were recorded. Collected lab examinations included blood routine test, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), synovial leukocyte counts, Receiver Operating Characteristic Curve-Area Under Curve (ROC-AUC) analysis will be used to evaluate the specific and sensitivity of diagnosis index. Moreover,microorganisms isolated from periprosthetic tissues and articular fluid. Postoperatively, the prosthesis was sent for ultrasound sonication. The sonicate extraction, implant surrounding tissue and synovium were sent for microbiologic culture, and the implant-surrounding-tissue were also sent for ...
In acoustic cavitation the spatial variation and time-dependent nature of the acoustic pressure field, whether it is a standing or propagating wave, together with the presence of other bubbles, particles and boundaries produces gradients and asymmetries in the flow field. This will inevitably lead to non-spherical bubble behaviour, often of short duration, before break-up into smaller bubbles which may act as nuclei for the generation of further bubbles. During the collapse phase, high temperatures and pressures will occur in the gaseous interior of the bubble. This paper concentrates on the non-spherical bubble extension to the earlier spherical-bubble studies for acoustic cavitation by exploiting the techniques that had previously been used to model incompressible hydraulic cavitation phenomena. Bubble behaviour near an oscillating boundary, jet impact and damage to boundaries, bubble interactions, bubble clouds and bubble behaviour near rough surfaces are considered. In many cases the key ...