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Invitrogen™ CellLight™ Nucleus-GFP, BacMam 2.0

Catalog No. C10602
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Catalog No. C10602 Supplier Invitrogen™ Supplier No. C10602
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Description

Provides easy way to label nuclei with green fluorescent protein in live cells

This ready-to-use construct is transfected into cells using BacMam 2.0 technology, where it expresses GFP fused to the SV40 nuclear localization sequence. You can observe nucleus-GFP behavior in live cells without the cellular toxicity associated with intercalators and label with multiple tracking or tracing dyes to image dynamic cellular processes.

Cells expressing CellLight constructs can also be fixed with formaldehyde for multiplexed imaging using immunocytochemical techniques.

BacMam Technology

CellLight Nucleus-GFP, BacMam 2.0, is a fusion construct of SV40 nuclear localization sequence and emGFP, providing accurate and specific targeting to cellular nucleus-GFP. This fusion construct is packaged in the insect virus baculovirus, which does not replicate in human cells and is designated as safe to use with biosafety level (BSL) 1 in most laboratories. BacMam technology ensures that most mammalian cell types are transduced/transfected with high efficiency and minimal toxicity. This transient transfection can be detected after overnight incubation for up to five days—enough time to carry out most dynamic cellular analyses. Like any transfection/transduction technique, the BacMam method does not transfect/transduce all of the cells with equal efficiency, making it poorly suited to cellular population studies or automated imaging/counting. CellLight reagents are ideal for experiments where cellular or subcellular co-locatization is required, or for cellular function studies that need special resolution.

  • Fast and convenient: simply add CellLight reagent to cells, incubate overnight, and image—or store frozen, assay-ready cells for later use
  • Highly efficient: up to 90% transduction of a wide range of mammalian cell lines, including primary cells, stem cells, and neurons
  • Flexible: co-transduce more than one BacMam reagent for multiplex experiments or co-localization studies; tightly control expression levels by simply varying the dose
  • Less toxic: CellLight reagents are non-replicating in mammalian cells and are suitable for biosafety level (BSL) 1 handling
  • Flow Cytometer Laser Lines: 488

Cell Analysis, Cell Structure, Cell Tracing & Tracking, Cellular Imaging, Immunocytochemistry (ICC), Immunofluorescence (IF), Nucleus, Nucleoli & Nuclear Envelope, Organelle Tracing

Order Info

Shipping Condition: Wet Ice

Specifications

Color Green
Content And Storage Store at 2°C to 6°C, protected from light. Do Not Freeze.
Excitation Wavelength Range 488⁄510
Dye Type GFP (EmGFP)
For Use With (Equipment) Confocal Microscope, Fluorescence Microscope
Quantity 1 mL
Detection Method Fluorescence
Emission Visible
Form Liquid
Product Line CellLight
Shipping Condition Wet Ice
Technique Fluorescence Intensity
Label Type Fluorescent Dye
Product Type Nucleus Label
SubCellular Localization Nucleus
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Frequently Asked Questions (FAQs)

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).
Is there any way to preserve the CellLights labeling beyond 5 days?

Cells transduced with the CellLights reagents can be stored frozen for several months after transduction, without loss of expression.
Are the CellLights products toxic to cells?

If the viral particles are used at the level we recommend, they are very well tolerated by cells.
For how long will the CellLights products label my cells?

The BacMam 2.0 CellLights typically express for 5 days after transduction.
What cell types can the CellLights products be used with?

The first generation BacMam reagents were shown to efficiently transduce over 90 cell types, including stable cell lines and primary cells. With BacMam 2.0, it is now possible to also efficiently transduce primary neurons and stem cells.
What safety precautions need to be taken to work with the BacMam reagent?

Our BacMam constructs do not replicate in mammalian cells, making them Biosafety Level I (the lowest biosafety level). They can be handled with the standard precautions used for any cell-based reagents.
I want to label the nuclei of live cells and track them over time. Can I use DAPI for this?

We do not recommend doing this. DAPI is considered to be a semi-permeant/impermeant nucleic acid stain. DAPI staining of live cells may be inconsistent. It is best used as a counterstain for fixed samples. Other cell permeable nucleic acid stains, such as Hoechst or the SYTO dyes may affect cellular function.

For mammalian cells, we recommend using the CellLight Nucleus transduction reagents, available in CFP, GFP and RFP. With these reagents, the cells are transduced overnight in a single labeling step and the next day the nuclei will fluoresce. The label may be retained for 3-5 days and should not affect cell function. Cytoplasmic cell tracking dyes such as the CellTracker dyes may also be used.

Do CellLight BacMam 2.0 reagents work on all cell types?

BacMam delivery technology works with over 90 mammalian cell types. However, cells with hematopoietic origin do not show effective transduction with CellLight BacMam 2.0 reagents.

I keep getting low transduction efficiency when using CellLight labeling reagents on my neurons. What can I do to improve the efficiency?

Neurons are more difficult to transduce than many other cells. The main way to improve transduction is to label with a higher number of particles per cell. For primary neurons, it can also help to transduce them at the time of plating rather than on established cultures. There can also be a slower onset of expression in neurons and peak expression often occurs on day 2-3 rather than 16 hours after transduction.
I want to track my cells with a nucleic acid stain, like DAPI or Hoechst dye. Do you recommend this?

This is not recommended. When these stains bind to DNA and RNA, they may affect the normal function of the nucleic acids, disrupting transcription, as well as replication. Other reagents, such as CellTracker dyes or Qtracker reagents are more optimized for tracking without disrupting normal activity. If a nuclear label is still desired, though, and the cells are mammalian and non-hematopoietic, CellLight nuclear reagents can transiently transfect cells to express GFP or RFP on a nuclear-expressing protein for up to several days without affecting function.

For Research Use Only. Not for use in diagnostic procedures.