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Invitrogen™ BacMam GFP Transduction Control (BacMam 2.0)

Catalog No. B10383
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Catalog No. B10383 Supplier Invitrogen™ Supplier No. B10383

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

Includes

BacMam GFP transduction control contains 1mL of virus at a concentration of approximately 1 x 108 viral particles/mL. Approximate fluorescence excitation/emission maxima for emGFP: 485/520 in nm.

Important part of the enhanced BacMam technology

The BacMam 2.0 Transduction Control lets you experience the power and convenience of enhanced BacMam technology, with the convenient readout of emerald GFP (emGFP); our Aequorea victoria emGFP is extremely bright and photostable and this non-targeted form is ideal for testing transduction. Check out how well BacMam 2.0 works with GFP in your model before advancing to the more specialized reagents. The BacMam technology is based on an insect virus (baculovirus) to help efficiently deliver and express genes in mammalian cells. The baculovirus has been modified to include an expression cassette for transgene expression in mammalian cells. BacMam 2.0 incorporate elements that help greatly enhance transduction efficiency and expression levels: a pseudotyped capsid protein for more efficient cell entry and genetic elements (enhanced CMV promoter and Woodchuck Post-transcriptional Regulatory Element), that boost expression levels. Baculoviruses do not replicate in mammalian cells and thus have an excellent safety profile and lack cytopathic effects on cells. Cell types previously not compatible with the technology (neuronal cells, T-cells), or only poorly transduced (some stem cells, CHO) are now transduced quantitatively in a simple, one step process.

  • Efficient: highly efficient (>90%) transduction of a wide range of mammalian cell lines, including primary cells, stem cells and neuronal cells
  • Fast and convenient: Simply add the BacMam 2.0 together with cells to your cover slip or microtiter plate, incubate overnight and assay or store frozen, assay-ready cells for later use
  • Safe: non-replicating in mammalian cells, lack of observable cytopathic effect and biosafety level (BSL) 1 handling
  • Enables Flexiblity: co-transduce more than one BacMam reagent for multiplex experiments or co-localization studies and tightly control expression levels by just varying the dose

Cell Analysis, Cell-Based Reporter Assays, Cellular Imaging, Fluorescent Protein Assays (GFP)

Order Info

Shipping Condition: Wet ice

Specifications

Color Green
Content And Storage The BacMam GFP ttransduction control contains 1 ml of virus at a concentration of approximately 1 × 10^8 viral particles⁄ml.
Approximate fluorescence excitation⁄emission maxima for emGFP: 485⁄520 in nm.
The BacMam transduction control should be stored at 4°C, protected from light. When stored as directed, it is stable for 6 months. DO NOT FREEZE.
Excitation Wavelength Range 485⁄520
Dye Type GFP (EmGFP)
For Use With (Equipment) Fluorescence Microscope, Fluorometer, HTS Reader, High Content Instrument, Flow Cytometer
Quantity 1 mL
Detection Method Fluorescence
Emission Visible
Form Liquid
Promoter CMV
Shipping Condition Wet Ice
Technique Fluorescence Intensity
Label Type Fluorescent Dye
Product Type BacMam GFP Transduction Control
Sub Cellular Localization Cytoplasm, 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.
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 am using your CellLight BacMam reagents for fluorescent protein-labeling of my cell organelles, but they aren't labeling well. How can I improve it?

First, make sure that your cells are mammalian and non-blood derived. If the cells have low transduction efficiency (i.e., only a few cells are labeling or are labeling only dimly), first try increasing the particle number per cell. The next thing to try is to use the BacMam Enhancer Solution. If, however, your cells show labeling, but the localization seems to be too generalized or ubiquitous within the cell, then you should reduce the particle number per cell. The manual gives a guideline and formula for calculating particle number per cell.

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