LentiBritePSD95-TagGFP2 LentiviralBiosen

LentiBritePSD95-TagGFP2 LentiviralBiosen

Stock Code: 3586361
Manufacturer Part No: 17-10227
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Application


Fluorescence microscopy imaging:
Primary rat hippocampal neuron cells were plated in a poly-D-lysine coated chambered cover glass for 5 days and transduced with lentiviral particles at an MOI of 20 for 24 hours. After media replacement and 2 weeks further incubation, cells were imaged live by oil immersion wide-field fluorescence microscopy. The PSD95-GFP displays a dotted distribution along the neurites.

Immunocytochemistry Comparison:
Rat hippocampal neuron cells were plated in a chamber slide and transduced with lentiviral particles at an MOI of 40 for 24 hours. After 24 hours, media was replaced and cells were then further incubated for 2 weeks. Immunocytochemical staining (red) of the same fields of view with a monoclonal antibody against PSD95 (Cat No. MAB1596) reveals similar expression patterns to the GFP-protein (green).

For optimal fluorescent visualization, it is recommended to analyze the target expression level within 24-48 hrs after transfection/infection for optimal live cell analysis, as fluorescent intensity may dim over time, especially in difficult-to-transfect cell lines. Infected cells may be frozen down after successful transfection/infection and thawed in culture to retain positive fluorescent expression beyond 24-48 hrs. Length and intensity of fluorescent expression varies between cell lines. Higher MOIs may be required for difficult-to-transfect cell lines.


Research CategoryNeuroscience


Research Sub CategorySynapse & Synaptic Biology


Components


TagPSD95-GFP2 Lentivirus:
One vial containing 25 µL of lentiviral particles at a minimum of 3 x 10E8 infectious units (IFU) per mL. For lot-specific titer information, please see “Viral Titer” in the datasheet.
Promoter
EF-1 (Elongation Factor-1)Multiplicty of Infection (MOI)
MOI = Ratio of # of infectious lentiviral particles (IFU) to # of cells being infected.
Typical MOI values for neurons to achieve high transduction efficiency and signal intensity are in the range of 20-40. For this target, some neuronal cell types may require higher or lower MOIs.
NOTE: MOI should be titrated and optimized by the end user for each cell type and lentiviral target to achieve desired transduction efficiency and signal intensity.


General description


Read our application note in Nature Methods!http://www.nature.com/app_notes/nmeth/2012/121007/pdf/an8620.pdf(Click Here!)Learn more about the advantages of our LentiBrite Lentiviral Biosensors! Click HereBiosensors can be used to detect the presence/absence of a particular protein as well as the subcellular location of that protein within the live state of a cell. Fluorescent tags are often desired as a means to visualize the protein of interest within a cell by either fluorescent microscopy or time-lapse video capture. Visualizing live cells without disruption allows researchers to observe cellular conditions in real time.Lentiviral vector systems are a popular research tool used to introduce gene products into cells

Quality Level100
ManufacturerSIGMA-ALDRICH
Technique(s)immunocytochemistry: suitable, cell based assay: suitable, transfection: suitable, single cell analysis: suitable, immunofluorescence: suitable

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