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Search results for: Mammalian Cell Extraction Kit500 assays

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#33282869   2020/11/17 To Up

Methods to Study Intracellular Movement and Localization of the Nucleotide Excision Repair Proteins at the DNA Lesions in Mammalian Cells.

Nucleotide excision repair (NER) is the most versatile DNA repair pathway that removes a wide variety of DNA lesions caused by different types of physical and chemical agents, such as ultraviolet radiation (UV), environmental carcinogen benzo[a]pyrene and anti-cancer drug carboplatin. The mammalian NER utilizes more than 30 proteins, in a multi-step process that begins with the lesion recognition within seconds of DNA damage to completion of repair after few hours to several days. The core proteins and their biochemical reactions are known from in vitro DNA repair assays using purified proteins, but challenge was to understand the dynamics of their rapid recruitment and departure from the lesion site and their coordination with other proteins and post-translational modifications to execute the sequential steps of repair. Here, we provide a brief overview of various techniques developed by different groups over last 20 years to overcome these challenges. However, more work is needed for a comprehensive knowledge of all aspects of mammalian NER. With this aim, here we provide detailed protocols of three simple yet innovative methods developed by many teams that range from local UVC irradiation to in situ extraction and sub-cellular fractionation that will permit study of endogenous as well as exogenous NER proteins in any cellular model. These methods do not require unique reagents or specialized instruments, and will allow many more laboratories to explore this repair pathway in different models. These techniques would reveal intracellular movement of these proteins to the DNA lesion site, their interactions with other proteins during repair and the effect of post-translational modifications on their functions. We also describe how these methods led us to identify hitherto unexpected role of poly(ADP-ribose) polymerase-1 (PARP1) in NER. Collectively these three simple techniques can provide an initial assessment of the functions of known and unknown proteins in the core or auxiliary events associated with mammalian NER. The results from these techniques could serve as a solid foundation and a justification for more detailed studies in NER using specialized reagents and more sophisticated tools. They can also be suitably modified to study other cellular processes beyond DNA repair.
Mihaela Robu, Rashmi G Shah, Girish M Shah

2379 related Products with: Methods to Study Intracellular Movement and Localization of the Nucleotide Excision Repair Proteins at the DNA Lesions in Mammalian Cells.

10501 kit10

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#33223826   2020/11/09 To Up

Multiscale Selectivity and in vivo Biodistribution of NRP-1Targeted Theranostic AGuIX Nanoparticles for PDT of Glioblastoma.

Local recurrences of glioblastoma (GBM) after heavy standard treatments remain frequent and lead to a poor prognostic. Major challenges are the infiltrative part of the tumor tissue which is the ultimate cause of recurrence. The therapeutic arsenal faces the difficulty of eradicating this infiltrating part of the tumor tissue while increasing the targeting of tumor and endogenous stromal cells such as angiogenic endothelial cells. In this aim, neuropilin-1 (NRP-1), a transmembrane receptor mainly overexpressed by endothelial cells of the tumor vascular system and associated with malignancy, proliferation and migration of GBM, highlighted to be a relevant molecular target to promote the anti-vascular effect of photodynamic therapy (VTP).
Mickaël Gries, Noémie Thomas, Joël Daouk, Paul Rocchi, Laurence Choulier, Justine Jubréaux, Julien Pierson, Aurélie Reinhard, Valérie Jouan-Hureaux, Alicia Chateau, Samir Acherar, Céline Frochot, François Lux, Olivier Tillement, Muriel Barberi-Heyob

1522 related Products with: Multiscale Selectivity and in vivo Biodistribution of NRP-1Targeted Theranostic AGuIX Nanoparticles for PDT of Glioblastoma.

5 G 5 G500 MG48 samples 100 G250 mg 1 G96 tests

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#33191941   2020/10/27 To Up

Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping.

Visceral adipose tissue (VAT) is an active metabolic organ composed mainly of mature adipocytes and stromal vascular fraction (SVF) cells, which release different bioactive molecules that control metabolic, hormonal, and immune processes; currently, it is unclear how these processes are regulated within the adipose tissue. Therefore, the development of methods evaluating the contribution of each cell population to the pathophysiology of adipose tissue is crucial. This protocol describes the isolation steps and provides the necessary troubleshooting guidelines for efficient isolation of viable mature adipocytes and SVF from human VAT biopsies in a single process, using a collagenase enzymatic digestion technique. Moreover, the protocol is also optimized to identify macrophage subsets and perform mature adipocyte RNA isolation for gene expression studies, which allows performing studies dissecting the interaction between these cell populations. Briefly, VAT biopsies are washed, minced mechanically, and digested to generate a single-cell suspension. After centrifugation, mature adipocytes are isolated by flotation from the SVF pellet. The RNA extraction protocol ensures a high yield of total RNA (including miRNAs) from adipocytes for downstream expression assays. Simultaneously, SVF cells are used to characterize macrophage subsets (pro- and anti-inflammatory phenotype) through flow cytometry analysis.
Guadalupe Estrada-Gutierrez, Eyerahi Bravo-Flores, Veronica Ortega-Castillo, Veronica Flores-Rueda, Ismael Mancilla-Herrera, Salvador Espino Y Sosa, Maribel Sánchez-Martínez, Otilia Perichart-Perera, Mario Solis-Paredes

1116 related Products with: Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage Phenotyping.

0.1ml (1mg/ml)0.1 mg1000 1 ml100ul200 1.0 mg100ug10 5mg

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#33178265   2020/10/28 To Up

Determination of the Cytotoxic Effect of Different Leaf Extracts from (Chrysobalanaceae).

Despite plants being a rich source of useful chemical compounds with different pharmacological properties, some of these compounds may be toxic to humans. , among its other important pharmacological activities, has been shown to have significant antiproliferative activity on cancer cell lines. Toxicity studies are required to determine the safety profile of in the consideration of its potential pharmaceutical benefits as a source of lead compounds in cancer therapy. The effects of on both the integrity of the erythrocyte membrane and on normal cells were determined. The dried leaf powder of was used in serial exhaustive extraction procedures using hexane, dichloromethane, ethyl acetate, acetone, ethanol, methanol, and water as solvents in addition to extraction using DCM: methanol in equal ratio. Alkaloids, flavonoids, and saponins were isolated from the ethanol extract. The leaf extracts were tested for haemolytic activity on sheep erythrocytes at concentrations of 0.625 to 5 mg/ml. The extracts were also tested for toxicity activity on normal mammalian cells such as the BALB/c mice peritoneal cells using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) at the concentrations of 6.3 to 50 g/ml. In the haemolysis assays, none of the plant extracts had a significant haemolytic activity with the saponin-enriched extract having the maximum haemolytic activity of 12.2% for a concentration of 5 mg/ml. In the MTT cell viability assay, none of the 11 plant extracts had significant cytotoxicity. The water extract, however, had significant ( < 0.01) proliferative activity towards the murine immune cells at all concentrations leaf extracts were, therefore, not toxic to both erythrocytes and immune cells, and the water extract may have immunostimulatory effects. It is concluded that leaf extracts are not toxic and, therefore, our results support the use of the plant for ethnomedicinal use.
Anesu Kundishora, Simbarashe Sithole, Stanley Mukanganyama

1303 related Products with: Determination of the Cytotoxic Effect of Different Leaf Extracts from (Chrysobalanaceae).

100tests 5 G100ug Lyophilized1 mg3x 500 ml100ug Lyophilized25 500 Units 500 ml 400Tests

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#33152052   2020/11/05 To Up

Toxicodendron vernicifluum Stokes extract inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c mice.

Toxicodendron vernicifluum Stokes has long been used as a food supplement and traditional herbal medicine in East Asia. We applied a new extraction method to produce Toxicodendron vernicifluum Stokes extract (TVSE), that doesn't contain urushiol (an allergenic toxin) but dose have higher levels of some flavonoids such as fustin and fisetin. This study was conducted to investigate the anticancer effects of TVSE in an in vivo system. Fifty BALB/c mice were acclimated for one week and then injected with 4T1 murine mammary carcinoma cells in mammary fat pads. After 7 days, the mice were randomly divided into 5 groups, and orally administered with 0, 50, 100, 200 or 400 mg of TVSE/kg body weight (BW)/day for 20 days. TVSE reduced tumor volume and weight dose-dependently. The expression of Ki67 was significantly reduced and the number of TUNEL-positive apoptotic cells was significantly increased in the TVSE-treated group over 100 mg/kg BW/day. While tumor nodules were not found in the liver, but only in lungs, the number of tumor nodules was reduced in a dose-dependent manner in the TVSE treated groups compared to the control group. In breast tumors, expression of platelet endothelial cell adhesion molecule (PECAM-1) and vascular endothelial growth factor (VEGF) was reduced by TVSE treatment. TVSE treatment significantly suppressed mRNA expression in tumors of matrix metalloproteinase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-1, urokinase-type plasminogen activator (uPA), intercellular adhesion molecule (ICAM)-1, and vascular cell adhesion molecule (VCAM)-1 while increasing plasminogen activator inhibitor (PAI)-1. These results suggest that TVSE is potentially beneficial for the suppression of breast cancer growth and its-associated lung metastasis.
Hyun Sook Lee, Jae In Jung, Kyeong-Hee Kim, Sang Jae Park, Eun Ji Kim

2637 related Products with: Toxicodendron vernicifluum Stokes extract inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c mice.

1.00 flask

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#33038776   2020/09/22 To Up

Characterization and hypoglycemic effects of sulfated polysaccharides derived from brown seaweed Undaria pinnatifida.

The brown seaweed Undaria pinnatifida polysaccharides show various biological activities, but their hypoglycemic activity and the underlying mechanism remain unclear. Here, three fractions of sulfated polysaccharides Up-3, Up-4, and Up-5 were prepared by microwave-assisted extraction from U. pinnatifida. In vitro assays demonstrated that Up-3 and Up-4 had strong α-glucosidase inhibitory activity, and Up-3, Up-4, and Up-5 could improve the glucose uptake in insulin-resistant HepG2 cells without affecting their viability. In vivo studies indicated Up-3 and Up-4 markedly reduced postprandial blood glucose levels. Up-U (a mixture of Up-3, Up-4, and Up-5), reduced fasting blood glucose levels, increased glucose tolerance and alleviated insulin resistance in HFD/STZ-induced hyperglycemic mice. Histopathological observation and hepatic glycogen measurement showed that Up-U alleviated the damage of the pancreas islet cell, reduced hepatic steatosis, and promoted hepatic glycogen synthesis. These findings suggest that Up-U could alleviate postprandial and HFD/STZ-induced hyperglycemia and was a potential agent for diabetes treatment.
Qi-Wu Zhong, Tao-Shun Zhou, Wen-Hui Qiu, Ya-Kun Wang, Qiao-Li Xu, Song-Ze Ke, Si-Jia Wang, Wei-Hua Jin, Jian-Wei Chen, Hua-Wei Zhang, Bin Wei, Hong Wang

2378 related Products with: Characterization and hypoglycemic effects of sulfated polysaccharides derived from brown seaweed Undaria pinnatifida.

100 mg 100 G 500 ml 1000 tests100ug5 mg50ug50 ug 5mg25 mg 100 G10 µg

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#32966064   2020/10/05 To Up

Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein.

There is an urgent need for robust and high-throughput methods for SARS-CoV-2 detection in suspected patient samples to facilitate disease management, surveillance, and control. Although nucleic acid detection methods such as reverse transcription polymerase chain reaction (RT-PCR) are the gold standard, during the current pandemic, the deployment of RT-PCR tests has been extremely slow, and key reagents such as PCR primers and RNA extraction kits are at critical shortages. Rapid point-of-care viral antigen detection methods have been previously employed for the diagnosis of respiratory viruses such as influenza and respiratory syncytial viruses. Therefore, the direct detection of SARS-CoV-2 viral antigens in patient samples could also be used for diagnosis of active infection, and alternative methodologies for specific and sensitive viral protein detection should be explored. Targeted mass spectrometry techniques have enabled the identification and quantitation of a defined subset of proteins/peptides at single amino acid resolution with attomole level sensitivity and high reproducibility. Herein, we report a targeted mass spectrometry assay for the detection of SARS-CoV-2 spike protein and nucleoprotein in a relevant biological matrix. Recombinant full-length spike protein and nucleoprotein were digested and proteotypic peptides were selected for parallel reaction monitoring (PRM) quantitation using a high-resolution Orbitrap instrument. A spectral library, which contained seven proteotypic peptides (four from spike protein and three from nucleoprotein) and the top three to four transitions, was generated and evaluated. From the original spectral library, we selected two best performing peptides for the final PRM assay. The assay was evaluated using mock test samples containing inactivated SARS-CoV-2 virions, added to in vitro derived mucus. The PRM assay provided a limit of detection of ∼200 attomoles and a limit of quantitation of ∼ 390 attomoles. Extrapolating from the test samples, the projected titer of virus particles necessary for the detection of SARS-CoV-2 spike and nucleoprotein detection was approximately 2 × 10 viral particles/mL, making it an attractive alternative to RT-PCR assays. Potentially, mass spectrometry-based methods for viral antigen detection may deliver higher throughput and could serve as a complementary diagnostic tool to RT-PCR. Furthermore, this assay could be used to evaluate the presence of SARS-CoV-2 in archived or recently collected biological fluids, in vitro-derived research materials, and wastewater samples.
Lisa H Cazares, Raghothama Chaerkady, Shao Huan Samuel Weng, Chelsea C Boo, Raffaello Cimbro, Hsiang-En Hsu, Sarav Rajan, William Dall'Acqua, Lori Clarke, Kuishu Ren, Patrick McTamney, Nicole Kallewaard-LeLay, Mahboobe Ghaedi, Yasuhiro Ikeda, Sonja Hess

1520 related Products with: Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein.

250tests96 reactions100 reactions100Tests100 100 µg250 mg100 reactions

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#32873806   2020/09/01 To Up

Representation of features as images with neighborhood dependencies for compatibility with convolutional neural networks.

Deep learning with Convolutional Neural Networks has shown great promise in image-based classification and enhancement but is often unsuitable for predictive modeling using features without spatial correlations. We present a feature representation approach termed REFINED (REpresentation of Features as Images with NEighborhood Dependencies) to arrange high-dimensional vectors in a compact image form conducible for CNN-based deep learning. We consider the similarities between features to generate a concise feature map in the form of a two-dimensional image by minimizing the pairwise distance values following a Bayesian Metric Multidimensional Scaling Approach. We hypothesize that this approach enables embedded feature extraction and, integrated with CNN-based deep learning, can boost the predictive accuracy. We illustrate the superior predictive capabilities of the proposed framework as compared to state-of-the-art methodologies in drug sensitivity prediction scenarios using synthetic datasets, drug chemical descriptors as predictors from NCI60, and both transcriptomic information and drug descriptors as predictors from GDSC.
Omid Bazgir, Ruibo Zhang, Saugato Rahman Dhruba, Raziur Rahman, Souparno Ghosh, Ranadip Pal

2597 related Products with: Representation of features as images with neighborhood dependencies for compatibility with convolutional neural networks.

100 assays96 Tests50 assays100Tests1,000 tests100 tests500 µl100 tests

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#32843049   2020/08/25 To Up

A novel two-step, direct-to-PCR method for virus detection off swabs using human coronavirus 229E.

Currently, one of the most reliable methods for viral infection detection are polymerase chain reaction (PCR) based assays. This process is time and resource heavy, requiring multiple steps of lysis, extraction, purification, and amplification procedures. Herein, we have developed a method to detect virus off swabs using solely shaker-mill based mechanical lysis and the transfer of the viral lysate directly to a PCR assay for virus detection, bypassing the substantial reagent and time investments required for extraction and purification steps.
Zachary P Morehouse, Caleb M Proctor, Gabriella L Ryan, Rodney J Nash

1023 related Products with: A novel two-step, direct-to-PCR method for virus detection off swabs using human coronavirus 229E.

96T100tests250tests0.1 mg96T100tests96 tests96tests 100ul0.05 mg96T20 ug

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