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).
Particles on the move: intracellular trafficking and asymmetric mitotic partitioning of nanoporous polymer particles.
Authors:Yan Y, Lai ZW, Goode RJ, Cui J, Bacic T, Kamphuis MM, Nice EC, Caruso F,
Journal:
PubMed ID:23713907
'Nanoporous polymer particles (NPPs) prepared by mesoporous silica templating show promise as a new class of versatile drug/gene delivery vehicles owning to their high payload capacity, functionality, and responsiveness. Understanding the cellular dynamics of such particles, including uptake, intracellular trafficking, and distribution, is an important requirement for their development as ... More
Advanced laboratory techniques for sample processing and immunolabeling using microwave radiation.
Authors:Ferris AM, Giberson RT, Sanders MA, Day JR,
Journal:J Neurosci Methods
PubMed ID:19520116
A better understanding of improved microwave technology has increased the benefits and versatility of the technique as it applies to all aspects of immunohistochemistry. The role of continuous magnetron power output (wattage) combined with precise control of sample heating demonstrated their significance to complex labeling protocols. Here, we present results ... More
Cytoskeletal control of CD36 diffusion promotes its receptor and signaling function.
The mechanisms that govern receptor coalescence into functional clusters--often a critical step in their stimulation by ligand--are poorly understood. We used single-molecule tracking to investigate the dynamics of CD36, a clustering-responsive receptor that mediates oxidized LDL uptake by macrophages. We found that CD36 motion in the membrane was spatially structured ... More
Spreading of neurodegenerative pathology via neuron-to-neuron transmission of ß-amyloid.
Alzheimer's disease (AD) is the major cause of dementia. During the development of AD, neurofibrillary tangles progress in a fixed pattern, starting in the transentorhinal cortex followed by the hippocampus and cortical areas. In contrast, the deposition of ß-amyloid (Aß) plaques, which are the other histological hallmark of AD, does ... More
Baculovirus-mediated gene transfer into mammalian cells.
Authors:Boyce FM, Bucher NL,
Journal:Proc Natl Acad Sci U S A
PubMed ID:8637876
This paper describes the use of the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a vector for gene delivery into mammalian cells. A modified AcMNPV virus was prepared that carried the Escherichia coli lacZ reporter gene under control of the Rous sarcoma virus promoter and mammalian RNA processing ... More