P3 Primary Cell 4D-Nucleofector® LV Kit XL
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Product Overview
The P3 Primary Cell 4D-NucleofectorTM LV Kit XL* is suited for larger scale transfection of primary cells in the LV NucleocuvetteTM Cartridge format when working with the 4D-NucleofectorTM LV Unit. With the LV NucleocuvetteTM Cartridge a variable volume of up to 20 mL can be transfected. The cartridge is automatically filled and emptied with help of the supplied input and output reservoirs. Alternatively bags can be used. An optional second input allows for separate feeding of mRNA to avoid its degradation.
This kit has been determined to be optimal for transfecting following primary cells and many others that use Solution P3 as optimal NucleofectorTM Solution:
- B cell (human)
- CD34+ cell (human)
- DC (human)
- ES (human), H9
- Macrophage (human)
- Monocyte (human)
- T cell (human), stim.
- T cell (mouse), unstim.
The respective cell type-specific Optimized Protocols or recommendations are available under "Instructions" or in our knowledge database. In these Optimized Protocols the best Nucleofection conditions are indicated. In addition, we share our experience and knowledge for treatment of individual cell types.
*Note: From the five primary cell solutions (P1 – P5) that come with the 4D-NucleofectorTM System only P3 is available for the large-scale formats so far. Solution P3 is suited for most immune and stem cells (e.g. T cells, dendritic cells, CD34 hematopoietic stem cells). For other solutions please contact our Scientific Support.
Benefits
- Closed Nucleofection of up to 1 billion primary cells
- Conditions transferable from existing smaller scale protocols
- Automatic processing via tubing system
- Transfection of up to 20 mL reaction volume in less than 10 minutes
Applications
- For large scale transfection of up to 1 x 109 cells
- Ex-vivo modification of human primary cells for the development and establishment of cell therapy applications (e.g. genome editing generation of CAR-T cells)
- Generation of large numbers of transiently modified primary cells for cell-based assays
Content & Storage
Content
22.5 mL P3 Primary Cell NucleofectorTM Solution
5 mL Supplement 1,
1 x LV NucleocuvetteTM Cartridge
2 x 4D-NucleofectorTM LV Reservoir
Instructions
Safety Data Sheets (SDS)
Certificate of Analysis (CoA)
Please enter Lot Number, including all zeros, located on the product label and please take into account that it is case sensitive.Brochures, White Papers etc.
Transfection of GFP mRNA into PBMCs
Fresh PBMCs (1x10⁸/mL) were transfected with GFP mRNA (20 µg/mL) using different Nucleofection® Vessels. For the fixed volume vessels (100 µL Nucleocuvette® Vessel and 1 mL Nucleocuvette® Cartridge), cells and mRNA for each sample were mixed separately, pipetted into the vessel and pulsed immediately. For the flow-through LV Nucleocuvette® Cartridge two feeding reservoirs were used to keep cells and mRNA separate as long as possible. Data are based on duplicates (experiment 1) or unicates (experiment 2).
