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	48-602MAG	Buffer Detection Kit for Magnetic Beads	1 Kit

17-10227 Sigma-AldrichLentiBrite™ PSD95-GFP Lentiviral Biosensor

LentiBrite™ PSD95-GFP Lentiviral Biosensor MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Key Applications: Transfection, Immunofluorescence, Immunocytochemistry, Track single cell and cell population migration (migration assay, cell culture)

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Przegląd

Replacement Information

Tabela kluczowych gatunków

Key Applications	Detection Methods
TFX, IF, ICC, Track single cell and cell population migration (migration assay, cell culture)	Fluorescent

Description
Catalogue Number	17-10227
Trade Name	LentiBrite Chemicon
Description	LentiBrite™ PSD95-GFP Lentiviral Biosensor
Overview	*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 Here Biosensors 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. Lentiviral transfection has advantages over non-viral methods such as chemical-based transfection including higher-efficiency transfection of dividing and non-dividing cells, long-term stable expression of the transgene, and low immunogenicity. EMD Millipore is introducing LentiBrite™ Lentiviral Biosensors, a new suite of pre-packaged lentiviral particles encoding important and foundational proteins of autophagy, apoptosis, and cell structure for visualization under different cell/disease states in live cell and in vitro analysis. Pre-packaged, fluorescently-tagged with GFP & RFP Higher efficiency transfection as compared to traditional chemical-based and other non-viral-based transfection methods Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types, such as primary cells or stem cells Non-disruptive towards cellular function EMD Millipore’s LentiBrite™ PSD-95-GFP lentiviral particles provide bright fluorescence and precise localization to enable live cell analysis of PSD dynamics in neuronal cells.
Background Information	The postsynaptic side of an excitatory synapse contains an electron- and protein-dense region, the postsynaptic density (PSD), that positions neurotransmitters adjacent to sites of presynaptic neurotransmitter release, and also concentrates signaling molecules. MAGUKs (membrane-associated guanylate kinases) serve both as central organizers of the PSD and as signaling molecules. One such MAGUK, PSD-95, is associated with the dendritic spines of mature synapses, and is relatively immobile compared to other MAGUKs. PSD-95 fused to fluorescent proteins has been employed to quantify turnover and mobility of PSD-95. EMD Millipore’s LentiBrite™ PSD-95-GFP lentiviral particles provide bright fluorescence and precise localization to enable live cell analysis of PSD dynamics in neuronal cells.

References

Product Information
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.
Detection method	Fluorescent
Quality Level	MQ100

Applications
Key Applications	Transfection Immunofluorescence Immunocytochemistry Track single cell and cell population migration (migration assay, cell culture)
Application Notes	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.

Applications

Key Applications

Transfection
Immunofluorescence
Immunocytochemistry
Track single cell and cell population migration (migration assay, cell culture)

Application Notes

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.

Biological Information
Gene Symbol	DLG4:PSD95
Purification Method	PEG precipitation
UniProt Number	P78352

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	Evaluated by transduction of HT-1080 cells and fluorescent imaging performed for assessment of transduction efficiency.
Usage Statement	This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges Merck to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form. Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Evaluated by transduction of HT-1080 cells and fluorescent imaging performed for assessment of transduction efficiency.

Usage Statement

This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges Merck to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Storage and Handling Lentivirus is stable for at least 4 months from date of receipt when stored at -80°C. After first thaw, place immediately on ice and freeze in working aliquots at -80°C. Frozen aliquots may be stored for at least 2 months. Further freeze/thaws may result in decreased virus titer and transduction efficiency. IMPORTANT SAFETY NOTE Replication-defective lentiviral vectors, such as the 3rd Generation vector provided in this product, are not known to cause any diseases in humans or animals. However, lentiviruses can integrate into the host cell genome and thus pose some risk of insertional mutagenesis. Material is a Risk Group 2 and should be handled under BSL2 controls. A detailed discussion of biosafety of lentiviral vectors is provided in Pauwels, K. et al. (2009). State-of-the-art lentiviral vectors for research use: Risk assessment and biosafety recommendations. Curr. Gene Ther. 9: 459-474.

Storage and Shipping Information

Storage Conditions

Storage and Handling
Lentivirus is stable for at least 4 months from date of receipt when stored at -80°C. After first thaw, place immediately on ice and freeze in working aliquots at -80°C. Frozen aliquots may be stored for at least 2 months. Further freeze/thaws may result in decreased virus titer and transduction efficiency.

IMPORTANT SAFETY NOTE
Replication-defective lentiviral vectors, such as the 3rd Generation vector provided in this product, are not known to cause any diseases in humans or animals. However, lentiviruses can integrate into the host cell genome and thus pose some risk of insertional mutagenesis. Material is a Risk Group 2 and should be handled under BSL2 controls. A detailed discussion of biosafety of lentiviral vectors is provided in Pauwels, K. et al. (2009). State-of-the-art lentiviral vectors for research use: Risk assessment and biosafety recommendations. Curr. Gene Ther. 9: 459-474.

Packaging Information
Material Size	1 vial (minimum of 3 x 10E8 IFU/mL)

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Numer katalogowy	GTIN
17-10227	04053252698064

Documentation

LentiBrite™ PSD95-GFP Lentiviral Biosensor MSDS

Title
Safety Data Sheet (SDS)

LentiBrite™ PSD95-GFP Lentiviral Biosensor Certificates of Analysis

Title	Lot Number
LentiBrite PSD95-GFP Lentiviral Biosensor - Q2043544	Q2043544
LentiBrite PSD95-GFP -2654749	2654749
LentiBrite PSD95-GFP -2716120	2716120
LentiBrite PSD95-GFP Lentiviral Biosensor - 3277558	3277558
LentiBrite PSD95-GFP Lentiviral Biosensor - 3700358	3700358
LentiBrite PSD95-GFP Lentiviral Biosensor - 3856156	3856156
LentiBrite PSD95-GFP Lentiviral Biosensor - 4128285	4128285
LentiBrite PSD95-GFP Lentiviral Biosensor - 4199647	4199647
LentiBrite PSD95-GFP Lentiviral Biosensor Packaged Lentiviral Particles	2945802
LentiBrite PSD95-GFP Lentiviral Biosensor Packaged Lentiviral Particles	3020582

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Kategorie

Life Science Research > Cell Analysis > Cell-based Assays > Lentiviral Biosensors

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