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Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography

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Evaluation of Anticancer activity of Silver Nanoparticles on the A549 Human Lung Carcinoma Cell Lines through Alamar Blue Assay

Authors: Nikita Sharma, Geeta Arya, R. Mankamna Kumari, Nidhi Gupta and Surendra Nimesh, date: 01/05/2019, view: 5415, Q&A: 0

[Abstract] Silver nanoparticles have been widely studied to possess antimicrobial as well as anticancer activity, and have found its applications in various fields including pharmaceutical industry, diagnostics, drug delivery, food industry, and others. For this purpose, several cell proliferation assays are widely used for the evaluation of anticancer ...

Monitoring Natural Killer Cell Function in Human Ovarian Cancer Cells of Ascitic Fluid

Authors: Vikash Kumar and Asok Mukhopadhyay, date: 12/20/2018, view: 2357, Q&A: 0

[Abstract] Natural killer (NK) cells are the major effectors of the innate immune system when activated resulting in modulation of immune response of the host defense through target cell lysis and secretion of cytokines. Precise functions of NK cells are essential for the treatment outcome of different virus infections and malignant diseases. NK cells impart ...

Detection of Cell Death in Planarians

Authors: Nídia de Sousa and Teresa Adell, date: 10/05/2018, view: 2954, Q&A: 0

[Abstract] Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the ...

An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist

Authors: Khalidah Ibraheem, Christopher J. Dunnill, Myria Ioannou, Albashir Mohamed, Balid Albarbar and Nikolaos T. Georgopoulos, date: 07/05/2018, view: 4460, Q&A: 0

[Abstract] One fundamental property of the TNR receptor (TNFR) family relates to how ‘signal quality’ (the extent of receptor ligation or cross-linking) influences the outcome of receptor ligation, for instance the induction of death in tumour cells. It is unequivocal that membrane-presented ligand (delivered to target cells via cell-surface presentation by ...

An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells

Authors: Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 4955, Q&A: 0

[Abstract] In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...

Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures

Authors: Jennifer Jung and Christian Behrends, date: 09/05/2017, view: 5232, Q&A: 0

[Abstract] Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy ...

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Authors: Sabine Pietkiewicz, Clara Wolfe, Jörn H. Buchbinder and Inna N. Lavrik, date: 01/05/2017, view: 6113, Q&A: 0

[Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 ...

In vitro Assays for the Detection of Calreticulin Exposure, ATP and HMGB1 Release upon Cell Death

Authors: Yuting Ma and Heng Yang, date: 12/20/2016, view: 12270, Q&A: 1

[Abstract] Accumulating evidence is revealing the essential role of immune system in cancer treatment. Certain chemotherapeutic drugs can potently induce the release of ‘cell death associated molecular patterns’ (CDAMPs), which accompanies cancer cell demise. CDAMPs can engage corresponding receptors on immune cells and stimulate immune responses to achieve ...

Determining the Influence of Small Molecules on Hypoxic Prostate Cancer Cell (DU-145) Viability Using Automated Cell Counting and a Cell Harvesting Protocol

Authors: John P Phelan, F Jerry Reen and Fergal O’Gara, date: 11/20/2016, view: 4789, Q&A: 0

[Abstract] Cell viability assays are an essential aspect of most cancer studies, however they usually require a considerable labor and time input. Here, instead of using the conventional microscopy and hemocytometer cell counting approach, we developed a cell harvesting protocol and combined it with the automated Countess Automated Cell Counter to generate ...

Phagocytosis Assay to Measure Uptake of Necroptotic Cancer Cells by BMDCs

Authors: Tania Løve Aaes, Dmitri V. Krysko and Peter Vandenabeele, date: 11/05/2016, view: 7751, Q&A: 0

[Abstract] This protocol is a flow cytometry-based method to measure the phagocytosis efficiency of necroptotic target cells by bone marrow-derived dendritic cells (BMDCs) in vitro (Aaes et al., 2016). The method is a slightly modified and updated version of the protocols used in previously published papers (Krysko et al., 2006; ...

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