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P3 Primary Cell 4D-Nucleofector® LV Kit L

Catalog #: V4LP-3002
For larger scale transfection of primary cells, e.g. human T cells or CD34+ hematopoietic stem cells in the 1 mL NucleocuvetteTM Cartridge format when working with the 4D-NucleofectorTM LV Unit
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Product Overview

The P3 Primary Cell 4D-NucleofectorTM LV Kit L is suited for larger scale transfection of primary cells in the 1 mL NucleocuvetteTM Cartridge format when working with the 4D-NucleofectorTM LV Unit. With the 1 mL NucleocuvetteTM Cartridge a fixed volume of 1 mL can be transfected. The cartridge is manually filled via a sterile injection port.

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 108 primary cells
  • Conditions transferable from existing smaller scale protocols
  • Simple manual filling via sterile injection port
  • Transfection within seconds

Applications

  • For large scale transfection of up to 1 x 108 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

2.25 mL P3 Primary Cell NucleofectorTM Solution

0.5 mL Supplement 1

50 µg pmaxGFPTM Vector (1 μg/μl in 10 mM Tris pH 8.0)

2 x 1 mL NucleocuvetteTM Cartridge

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.

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).

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