MilliporeSigma
All Photos(4)

Documents

17-10387

Sigma-Aldrich

LentiBrite GFP Control Lentiviral Biosensor

Sign Into View Organizational & Contract Pricing

Synonym(s):
Lentiviral biosensor control particles
eCl@ss:
34360190

manufacturer/tradename

Chemicon®
LentiBrite

Quality Level

technique(s)

cell based assay: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
transfection: suitable

detection method

fluorometric

shipped in

dry ice

Related Categories

Compare Similar Items

View Full Comparison

Show Differences

1 of 4

This Item
17-1040917-1022917-10208
Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

manufacturer/tradename

Chemicon®

manufacturer/tradename

Chemicon®, LentiBrite

manufacturer/tradename

Chemicon®, LentiBrite

manufacturer/tradename

Chemicon®, LentiBrite

detection method

fluorometric

detection method

fluorometric

detection method

fluorometric

detection method

fluorometric

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 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 GFP Control lentiviral particles provide bright fluorescence to enable live-cell analysis in difficult-to-transfect cell types.
The use of genetically encoded fluorescent proteins has revolutionized cell biology by enabling real-time imaging of cells and subcellular structures. A diverse palette of fluorescent proteins is available, but these proteins vary in their suitability for live-cell imaging. EMD Millipore’s LentiBriteTM GFP Control Lentiviral Biosensor employs TagGFP2, the improved variant of the Aequorea macrodactyla GFP-like protein. TagGFP2 exhibits bright green fluorescence comparable to that of EGFP, with excitation/emission maxima at 483 and 506 nm, respectively. TagGFP2 is specially optimized for expression at 37°C and is therefore well-suited for use in mammalian cells. EMD Millipore’s LentiBrite GFP Control lentiviral particles provide bright fluorescence to enable live-cell analysis in difficult-to-transfect cell types.

application

Fluorescence Microscopy
Imaging:
(See Figure 1 in datasheet)
HeLa cells were plated in a chamber slide and transduced with lentiviral particles at an MOI of 40 for 24 hours. After media replacement and 48 hours further incubation, cells were fixed with formaldehyde and mounted. Images were obtained by oil immersion wide-field fluorescence microscopy. The GFP displays a uniform green fluorescent distribution in the cells.


Immunocytochemistry Comparison:
(See Figure 2 in datasheet)
Similar to Figure 1 (see datasheet), U2OS 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 48 hours.

Hard-to-transfect Cell Types:
(See Figure 3 in datasheet)
Primary cell types HUVEC or HuMSC were plated in chamber slides and transduced with lentiviral particles at an MOI of 40 for 24 hours.

Additional cell type:
(See Figure 4 in datasheet)
Primary Rat Cortical Neuron cells were treated as in Figure 1 (see datasheet).

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 Category
Cell Structure
Research Sub Category
All

Components

TagGFP2 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 lot specific “Viral Titer” in the product specifications of 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 high transduction efficiency and signal intensity are in the range of 20-40. For this target, some cell types may require lower MOIs (e.g., HT-1080, HeLa, U2OS, human mesenchymal stem cells (HuMSC)), while others may require higher MOIs (e.g., human umbilical vein endothelial cells (HUVEC)).
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.

Quality

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

Physical form

PEG precipitation

Storage and Stability

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.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

Already Own This Product?

Documents related to the products that you have purchased in the past have been gathered in the Document Library for your convenience.

Visit the Document Library

Difficulty Finding Your Product Or Lot/Batch Number?

Product numbers are combined with Pack Sizes/Quantity when displayed on the website (example: T1503-25G). Please make sure you enter ONLY the product number in the Product Number field (example: T1503).

Example:

T1503
Product Number
-
25G
Pack Size/Quantity

Additional examples:

705578-5MG-PW

PL860-CGA/SHF-1EA

MMYOMAG-74K-13

1000309185

enter as 1.000309185)

Having trouble? Feel free to contact Technical Service for assistance.

Lot and Batch Numbers can be found on a product's label following the words 'Lot' or 'Batch'.

Aldrich Products

  • For a lot number such as TO09019TO, enter it as 09019TO (without the first two letters 'TO').

  • For a lot number with a filling-code such as 05427ES-021, enter it as 05427ES (without the filling-code '-021').

  • For a lot number with a filling-code such as STBB0728K9, enter it as STBB0728 without the filling-code 'K9'.

Not Finding What You Are Looking For?

In some cases, a COA may not be available online. If your search was unable to find the COA you can request one.

Request COA

Related Content

Fluorescent lentiviral particles encoding important GFP/RFP fusion proteins related to autophagy, apoptosis, and cell structure that enables live cell imaging.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service