Search results for: Cytosol/Particulate Separation Kit
#28841990 2017/08/26 Save this To Up
High-precision quantitation of a tuberculosis vaccine antigen with capillary-gel electrophoresis using an injection standard.Analysis of proteinogenic vaccine antigens in a quality control environment requires an accurate, precise, and reliable method for protein separation and quantitation. While having multiple advantages over the classical SDS-PAGE, capillary gel electrophoresis (CGE) has not yet become a standard tool in vaccine antigen analysis. Here we report on development of a CGE-based method for quantitative analysis of a tuberculosis vaccine fusion antigen protein, H4, currently in clinical trials. We demonstrate that our method can monitor antigen purity and relative quantity with greater precision and accuracy versus SDS-PAGE. In addition, due to use of direct light-absorbance detection, the CGE method is suitable for absolute quantitation, an application for which SDS-PAGE is limited due to the need for staining and limited dynamic range of detection. To further improve the performance of our quantitation method, we introduced Bovine Serum Albumin (BSA) as an injection standard to correct for signal variance associated with the injected sample volume. We found that, for our specific application, BSA was more appropriate as an injection standard versus one provided in a commercial kit, in terms of precision and accuracy for quantitation of H4. In addition to providing better method performance versus SDS-PAGE, CGE is also faster and less resource-intensive. We conclude that CGE should be considered as a replacement for traditional SDS-PAGE methods for vaccine antigen quantitation in a quality-control environment.
1536 related Products with: High-precision quantitation of a tuberculosis vaccine antigen with capillary-gel electrophoresis using an injection standard.HBV surface recombinant a Mycobacterium tuberculosi Rabbit Polyclonal to Myco MAP-2 antigen alpha Tubulin TUBA1A TUB Hepatitis B Core Antigen Hepatitis B Core Antigen Hepatitis B Core Antigen Hepatitis B Core Antigen Epithelial Membrane Anti Epithelial Membrane Anti CD30, Ki-1 Antigen; Clon
#28811118 2017/08/16 Save this To Up
Prediction and characterization of the stability enhancing effect of the Cherry-Tag™ in highly concentrated protein solutions by complex rheological measurements and MD simulations.Solution stability attributes are one of the key parameters within the production and launching phase of new biopharmaceuticals. Instabilities of active biological compounds can reduce the yield of biopharmaceutical productions, and may induce undesired reactions in patients, such as immunogenic rejections. Protein solution stability thus needs to be engineered and monitored throughout production and storage. In contrast to the gold standard of long-term storage experiments applied in industry, novel experimental and in silico molecular dynamics tools for predicting protein solution stability can be applied within several minutes or hours. Here, a rheological approach in combination with molecular dynamics simulations are presented, for determining and predicting long-term phase behavior of highly concentrated protein solutions. A diversity of liquid phase conditions, including salt type, ionic strength, pH and protein concentration are tested in a Glutathione-S-Transferase (GST) case study, in combination with the enzyme with and without solubility-enhancing Cherry-Tag™. The rheological characterization of GST and Cherry-GST solutions enabled a fast and efficient prediction of protein instabilities without the need of long-term protein phase diagrams. Finally, the strong solubility enhancing properties of the Cherry-Tag™ were revealed by investigating protein surface properties in MD simulations. The tag highly altered the overall surface charge and hydrophobicity of GST, making it less accessible to alteration by the chemical surrounding.
2725 related Products with: Prediction and characterization of the stability enhancing effect of the Cherry-Tag™ in highly concentrated protein solutions by complex rheological measurements and MD simulations.Recombinant Human PKC the Recombinant Human PKC the Recombinant Human PKC the Apoptosis Phospho-Specifi EGF Phospho-Specific Arra Thermal Shaker with cooli TP53 & MDM2 Protein Prote CDKN1A & MDM2 Protein Pro AKT1 & MDM2 Protein Prote IGF1R & MDM2 Protein Prot CASP3 & MDM2 Protein Prot Rabbit Anti-Rat Androgen
#28803649 2017/08/14 Save this To Up
Root cause investigation of deviations in protein chromatography based on mechanistic models and artificial neural networks.In protein chromatography, process variations, such as aging of column or process errors, can result in deviations of the product and impurity levels. Consequently, the process performance described by purity, yield, or production rate may decrease. Based on visual inspection of the UV signal, it is hard to identify the source of the error and almost unfeasible to determine the quantity of deviation. The problem becomes even more pronounced, if multiple root causes of the deviation are interconnected and lead to an observable deviation. In the presented work, a novel method based on the combination of mechanistic chromatography models and the artificial neural networks is suggested to solve this problem. In a case study using a model protein mixture, the determination of deviations in column capacity and elution gradient length was shown. Maximal errors of 1.5% and 4.90% for the prediction of deviation in column capacity and elution gradient length respectively demonstrated the capability of this method for root cause investigation.
1238 related Products with: Root cause investigation of deviations in protein chromatography based on mechanistic models and artificial neural networks.HIV 1 intergase antigen. 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 Native Influenza HA (A Br
#28800539 2017/08/11 Save this To Up
Development of a rapid and sensitive multiple reaction monitoring proteomic approach for quantification of transporters in human liver tissue.With increasing knowledge on the role of hepatic transporters in drug disposition, numerous efforts have been described to quantify the expression of human hepatic transporters. However, reported quantitative proteomic approaches often require long analysis times. Additionally, greater assay sensitivity is still necessary for less abundant transporters or limited quantity of samples (e.g. hepatocytes and liver tissue). In the present study, an LC-MS/MS method for rapid and simultaneous quantification of 12 hepatic transporters (BCRP, BSEP, MATE1, MRP2, MRP3, MRP4, NTCP, OATP1B1, 1B3, 2B1, OCT1, and P-gp) was developed. Using a high LC flow rate (1.5mL/min) and fast LC gradient (4min total cycle time), the run time was markedly reduced to 4min, which was much shorter than most previously published assays. Chromatographic separation was achieved using ACE UltraCore SuperC18 50mm×2.1mm 5-μm HPLC column. In addition, greater analytical sensitivity was achieved with both high LC flow rate/fast LC gradient and post-column infusion of ethylene glycol. The on-column LLOQ for signature peptides in this method ranged from 0.194 to 0.846 femtomoles. The impact of five protein solubilizers, including extraction buffer II of ProteoExtract Native Membrane Protein Extraction Kit, 3% (w/v) sodium deoxycholate, 20% (v/v) Invitrosol, 0.2% (w/v) RapiGest SF, and 10% (w/v) formamide on total membrane protein extraction and trypsin digestion was investigated. Sodium deoxycholate was chosen because of good total membrane protein extraction and trypsin digestion efficiency, as well as no significant MS interference. Good precision (within 15% coefficient of variation) and accuracy (within ±15% bias), and inter-day trypsin digestion efficiency (within 28% coefficient of variation) was observed for quality controls. This method can quantify human hepatic transporter expression in a high-throughput manner and due to the increased sensitivity can be used to investigate the down-regulation of hepatic transporter protein (e.g., different ethnic groups and liver disease patients).
1436 related Products with: Development of a rapid and sensitive multiple reaction monitoring proteomic approach for quantification of transporters in human liver tissue.Multiple organ tumor tiss Multiple organ tumor tiss Multiple organ tumor tiss Frozen multiple human org Multiple lung carcinoma ( Multiple diseases of live Inflammation (Human) Quan Inflammation (Human) Quan Inflammation (Human) Quan Multiple organs tumor and Multiple organ cancer tis Liver disease spectrum ti
#28766790 2017/08/02 Save this To Up
Spatial and Lateral Control of Functionality by Rigid Molecular Platforms.Surface mounted molecular devices have received significant attention in the scientific community because of their unique ability to construct functional materials. The key involves the platform on which the molecular device works on solid substrates, such as in solid-liquid or solid-vacuum interfaces. Here, we outline the concept of rigid molecular platforms to immobilize active functionality atop flat surfaces in a controllable manner. Most of these (multipodal) platforms have at least three anchoring groups to control the spatial arrangement of the protruding functional moieties and form mechanically stable and electronically tuned contacts to the underlying substrate. Another approach is based on employing of flat aromatic scaffolds bearing perpendicular functionalities that form stable lateral assemblies on various surfaces. Emphasis is placed on the need for controllable assembly and separation of these tailor-made molecules that expose functionalities at the molecular scale. The discussions are focused on the different molecular designs realizing functional 3D architectures on surfaces, the role of various anchoring strategies to control the spatial arrangement, and structural considerations controlling physical features like the coupling to the surface or the available space for sterically demanding molecular operations.
2200 related Products with: Spatial and Lateral Control of Functionality by Rigid Molecular Platforms.EZH2 KMT6 Control Peptid GFP control peptide anti GFP Control Peptide Cytokeratin, High Molecu Cytokeratin, High Molecu Cytokeratin, High Molecu Cytokeratin, High Molecu Cytokeratin, High Molecu Cytokeratin, High Molecu Growth Differentiation Fa Androgen Receptor (Phosph Androgen Receptor (Phosph
#28758288 2017/07/31 Save this To Up
Stimuli-Responsive Metal-Organic Frameworks with Photoswitchable Azobenzene Side Groups.Metal-organic frameworks (MOFs) are nanoporous, crystalline hybrid materials, which enable various functionalities by incorporating functional organic molecules. By using organic linker molecules that possess photoswitchable azobenzene side groups, the remote control over certain properties was introduced to MOFs. Different MOF materials in the form of powders and thin films have been used to demonstrate the photoswitching. The applications of these stimuli-responsive nanoporous solids range from switching the adsorption capacity of various gases over remote-controlled release of guest molecules to continuously tunable membrane separation of molecular mixtures. A particular focus of this review is the effect of the azobenzene photoswitching on the host-guest interaction, enabling smart applications of the material. Steric hindrance, which may suppress the photoswitching in some MOF structures, is also discussed.
2327 related Products with: Stimuli-Responsive Metal-Organic Frameworks with Photoswitchable Azobenzene Side Groups.Azobenzene 2 sulfenyl bro Azobenzene C12H10N2 CAS: Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra Rabbit Anti-Metal ion tra
#28749520 2017/07/27 Save this To Up
Photocurrent spectroscopy of dye-sensitized carbon nanotubes.Monochiral (7,5) single walled carbon nanotubes (SWCNTs) are integrated into a field effect transistor device in which the built-in electric field at the nanotube/metal contact allows for exciton separation under illumination. Variable wavelength spectroscopy and 2D surface mapping of devices consisting of 10-20 nanotubes are performed in the visible region and a strong correlation between the nanotube's second optical transition (S22) and the photocurrent is found. After integration, the SWCNTs are non-covalently modified with three different fluorescent dye molecules with off-resonant absorption maxima at 532 nm, 565 nm, and 610 nm. The dyes extend the absorption properties of the nanotube and contribute to the photocurrent. This approach holds promise for the development of photo-detectors and for applications in photovoltaics and biosensing.
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#28738961 2017/07/25 Save this To Up
[Circular RNA CircHIPK3 Promotes NCI-H1299 and NCI-H2170 Cell Proliferation through miR-379 and its Target IGF1].It has been proven that the circular RNA, possessing a stable covalently closed continuous loop, is a type of RNA molecule which is expressed widespread in mammals. The circular RNA circHIPK3 is abundantly expressed in hepatocellular carcinoma (HCC) and promotes tumourgenesis. However, a role for circHIPK3 has not been systematically examined in non-small cell lung cancer (NSCLC). In this study, we investigated whether circHIPK3 has an effect on cell proliferation in the NSCLC cell lines NCI-H1299 and NCI-H2170 and the underlying molecular mechanisms.
1375 related Products with: [Circular RNA CircHIPK3 Promotes NCI-H1299 and NCI-H2170 Cell Proliferation through miR-379 and its Target IGF1].Epidermal Growth Factor ( Epidermal Growth Factor ( T-cell proliferation grad TCCI T cell proliferation TCCI T cell proliferation T-cell proliferation grad TCBI T cell proliferation TCBI T cell proliferation T-cell proliferation grad TCPI T cell proliferation TCPI T cell proliferation T-cell proliferation grad
#28731571 2017/07/21 Save this To Up
Model-Based Investigation on the Mass Transfer and Adsorption Mechanisms of Mono-Pegylated Lysozyme in Ion-Exchange Chromatography.Recent studies highlighted the potential of PEGylated proteins to improve stabilities and pharmacokinetics of protein drugs. Ion-exchange chromatography (IEX) is among the most frequently used purification methods for PEGylated proteins. However, the underlying physical mechanisms allowing for a separation of different PEGamers (proteins with a varying number of attached PEG molecules) are not yet fully understood. In this work, mechanistic chromatography modeling is applied to gain a deeper understanding of the mass transfer and adsorption/desorption mechanisms of mono-PEGylated proteins in IEX. Using a combination of the general rate model (GRM) and the steric mass action (SMA) isotherm, simulation results in good agreement with the experimental data are achieved. During linear gradient elution of proteins attached with PEG of different molecular weight, similar peak heights, and peak shapes at constant gradient length are observed. A superimposed effect of increased desorption rate and reduced diffusion rate as a function of the hydrodynamic radius of PEGylated proteins is identified to be the reason of this anomaly. That is why the concept of the diffusion-desorption-compensation effect is proposed. In addition to the altered elution orders, PEGylation results in a considerable decrease of maximum binding capacity. By using the SMA model in a kinetic formulation, the adsorption behavior of PEGylated proteins in the highly concentrated state is described mechanistically. An exponential increase in the steric hindrance effect with increasing PEG molecular weight is observed. This suggests the formation of multiple PEG layers in the interstitial space between bound proteins and an associated shielding of ligands on the adsorber surface to be the cause of the reduced maximum binding capacity. The presented in silico approach thus complements the hitherto proposed theories on the binding mechanisms of PEGylated proteins in IEX.
2818 related Products with: Model-Based Investigation on the Mass Transfer and Adsorption Mechanisms of Mono-Pegylated Lysozyme in Ion-Exchange Chromatography.ABT-263 Mechanisms: Bcl-2 ABT-737 Mechanisms: Bcl-2 AS-703026 Mechanisms: MEK AZD-6244 Mechanisms: MEK AZD-8055 Mechanisms: mTOR BEZ-235 Mechanisms: PI3K BMS-754807 Mechanisms: IG Bortezomib (PS-341) Mecha GDC-0941 Mechanisms: PI3K INK-128 Mechanisms: mTORC LBH-589 (Panobinostat) Me PD-0332991 (Palbociclib)
#28726276 2017/07/20 Save this To Up
The colonic groove of the plains viscacha (Lagostomus maximus): Histochemical evidence of an abrupt change in the glycosylation pattern of goblet cells.The ascending colon of most rodent species shows a longitudinal colonic groove that works as a retrograde transport pathway for a mixture of bacteria and mucus toward the cecum. We describe the morphology and glycosylation pattern of the colonic groove of Lagostomus maximus to analyze the role of mucins in this anatomical feature. We also studied the distribution pattern of the interstitial cells of Cajal (ICC) to evaluate their regulatory influence on gut motility. The groove originated near the cecocolic junction and extended along the mesenteric side of the ascending colon, limited at both ends by nonpapillated ridges. These ridges divided the lumen of the ascending colon into two compartments: a narrow channel and a large channel, called the groove lumen and the main lumen, respectively. The histochemical analysis showed differences in the glycosylation pattern of the goblet cells inside and outside the groove. Unlike the mucosa lining the main lumen of the colon, the groove was rich in goblet cells that secrete sulfomucins. The PA/Bh/KOH/PAS technique evidenced an abrupt change in the histochemical profile of goblet cells, which presented a negative reaction in the groove and a strongly positive one in the rest of the colonic mucosa. The anti-c-kit immunohistochemical analysis showed different ICC subpopulations in the ascending colon of L. maximus. Of all types identified, the ICC-SM were the only cells located solely within the colonic groove.
2303 related Products with: The colonic groove of the plains viscacha (Lagostomus maximus): Histochemical evidence of an abrupt change in the glycosylation pattern of goblet cells.FDA Standard Frozen Tissu FDA Standard Frozen Tissu FDA Standard Frozen Tissu FDA Standard Frozen Tissu FDA Standard Frozen Tissu FDA Standard Frozen Tissu Multiple organ tumor tiss TCP-1 theta antibody Sour Rabbit anti PKC theta (Ab Rabbit anti PKC theta (Ab Rabbit anti PKC theta (Ab Thermal Shaker with cooli
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