How can I increase the transduction efficiency with the BacMam 2.0 reagents such as the the CellLight and Premo products?
Try varying particle-to-cell ratio (PPC), incubation volume, temperature and, cell density (if adherent cells are transduced). For adherent cells, we recommend a confluence of about 70%. Following the PPC, adjusting the volume is the next best parameter to change to optimize protein expression. If that doesn't work, you can also use the BacMam Enhancer Kit (Cat. No. B10107).
Faecal excretion of ciprofloxacin after a single oral dose and its effect on faecal bacteria in healthy volunteers.
Authors:Pecquet S, Ravoire S, Andremont A,
Journal:J Antimicrob Chemother
PubMed ID:2211433
'High concentrations of ciprofloxacin have been shown to persist in the faeces of volunteers for several days after a week of oral treatment with this drug, which was also found to have a prolonged effect on aerobic Gram-negative intestinal bacteria. To determine whether a shorter course of ciprofloxacin would have ... More
Intracellular trafficking mechanism, from intracellular uptake to extracellular efflux, for phospholipid/cholesterol liposomes.
Authors:Un K, Sakai-Kato K, Oshima Y, Kawanishi T, Okuda H,
Journal:Biomaterials
PubMed ID:22858002
'Liposomes are widely used as drug delivery vehicles to transfer chemotherapeutic agents, proteins, and nucleic acids into target cells. To improve therapeutic effects and reduce unexpected toxic side-effects, it is necessary to understand the mechanism of liposomal uptake into cells, and the intracellular fate of internalized liposomes. The intracellular fate ... More
Utilization of fluorescent probes for the quantification and identification of subcellular proteomes and biological processes regulated by lipid peroxidation products.
Authors:Cummins TD, Higdon AN, Kramer PA, Chacko BK, Riggs DW, Salabei JK, Dell'italia LJ, Zhang J, Darley-Usmar VM, Hill BG,
Journal:Free Radic Biol Med
PubMed ID:22954622
Oxidative modifications to cellular proteins are critical in mediating redox-sensitive processes such as autophagy, the antioxidant response, and apoptosis. The proteins that become modified by reactive species are often compartmentalized to specific organelles or regions of the cell. Here, we detail protocols for identifying the subcellular protein targets of lipid ... More
Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells.
Phase imaging with a high-resolution wavefront sensor is considered. This is based on a quadriwave lateral shearing interferometer mounted on a non-modified transmission white-light microscope. The measurement technology is explained both in the scope of wave optics and geometrical optics in order to discuss its implementation on a conventional microscope. ... More
Combination of Cl-IB-MECA with paclitaxel is a highly effective cytotoxic therapy causing mTOR-dependent autophagy and mitotic catastrophe on human melanoma cells.
Authors:Soares AS, Costa VM, Diniz C, Fresco P,
Journal:
PubMed ID:24659394
Metastatic melanoma is the deadliest form of skin cancer. It is highly resistant to conventional therapies,particularly to drugs that cause apoptosis as the main anticancer mechanism. Recently, induction of autophagic cell death is emerging as a novel therapeutic target for apoptotic-resistant cancers. We aimed to investigate the underlying mechanisms elicited ... More