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For mammalian fibroblasts lacking a cell-type specific Amaxa™ Optimized Protocol, we offer a selection of kits that can be used to easily define optimal Nucleofection™ Conditions. The P2 and P3 Primary Cell Kits are suited for optimizations of mammalian fibroblasts on the 4D-Nucleofector™ and the 96-well Shuttle™ System. A cell group-specific Basic Kit is suited for optimization of mammalian fibroblasts using the Nucleofector™ II/2b Device.
Knockdown of N-cadherin by Nucleofection™ of primary human colon myofibroblasts with siRNA
Western blot (left) showing the knockdown of N-cadherin upon Nucleofection™ of myofibroblasts with combinations of specific siRNAs derived from different regions of N-cadherin cDNA (siN-cad 2+3 or 3+4) or control siRNA (con). Cadherin-11 expression is unchanged. Tubulin was used as loading control. (right) Inhibiting effect of N-cadherin knockdown on TGF-b-stimulated invasion of myofibroblasts in a spheroidcell-collagen-invasion-assay after 2 and 4 days. (Data reproduced from De Wever et al. (2004) J Cell Sci 117(Pt 20), 4691-703 with permission of the Company of Biologists and of the authors.)
For the 4D-Nucleofector™ Kits "L" and "S" indicate the Nucleocuvette™ Vessel format. L = 100 µl single Nucleocuvette™, S = 16-well Nucleocuvette™ Strip.