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).
'Monocarboxylate transporter 8 (MCT8) is a thyroid hormone (TH)-specific transporter. Mutations in the MCT8 gene are associated with Allan-Herndon-Dudley Syndrome (AHDS), consisting of severe psychomotor retardation and disturbed TH parameters. To study the functional consequences of different MCT8 mutations in detail, we combined functional analysis in different cell types with ... More
Single-cell redox imaging demonstrates a distinctive response of dopaminergic neurons to oxidative insults.
Authors:Horowitz MP, Milanese C, Di Maio R, Hu X, Montero LM, Sanders LH, Tapias V, Sepe S, van Cappellen WA, Burton EA, Greenamyre JT, Mastroberardino PG,
Journal:Antioxid Redox Signal
PubMed ID:21395478
'The study of the intracellular oxido-reductive (redox) state is of extreme relevance to the dopamine (DA) neurons of the substantia nigra pars compacta. These cells possess a distinct physiology intrinsically associated with elevated reactive oxygen species production, and they selectively degenerate in Parkinson''s disease under oxidative stress conditions. To test ... 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
BPIFB3 regulates autophagy and coxsackievirus B replication through a noncanonical pathway independent of the core initiation machinery.
Authors:Delorme-Axford E, Morosky S, Bomberger J, Stolz DB, Jackson WT, Coyne CB,
Journal:
PubMed ID:25491355
'Enteroviruses require autophagy to facilitate the formation of autophagosome (AP)-like double-membrane vesicles that provide the scaffolding for RNA replication. Here, we identify bactericidal/permeability-increasing protein (BPI) fold-containing family B, member 3 (BPIFB3) as a gene whose silencing greatly enhances coxsackievirus B (CVB) replication and induces dramatic alterations in the morphology of ... More
The amyloid precursor protein copper binding domain histidine residues 149 and 151 mediate APP stability and metabolism.
One of the key pathological hallmarks of Alzheimer disease (AD) is the accumulation of the APP-derived amyloid ß peptide (Aß) in the brain. Altered copper homeostasis has also been reported in AD patients and is thought to increase oxidative stress and to contribute to toxic Aß accumulation and regulate APP ... More