The Amaxa Cell Line Nucleo- factor Kit R (L-013 program; Lonza) was used to electroporate 5 × 106 IIA1.6 B Lymphoma cells for different transfections. For Centrin-GFP plasmid transfection we used 2 µg of DNA, and cells were cultured for 16 h before functional analysis. In the case of transfection with siRNA we used Silencer® Select (Ambion/Thermofisher) against Nesprin-1(Syne1): s234287 (#1), s234288 (#2); siRNA Sun1: s94911 (#1), s94912 cells (#2), these were incubated for 48 h before analysis. As a control, we used a scrambled siRNA (Qiagen) at 10nM.
Amaxa cell line nucleo factor kit r
Manufactured by Lonza
The Amaxa Cell Line Nucleofector Kit R is a laboratory equipment product designed for the transfection of cell lines. It provides a standardized protocol for the efficient delivery of nucleic acids into a variety of cultured cell types.
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3 protocols using amaxa cell line nucleo factor kit r
1
Electroporation and siRNA Transfection for B Lymphoma
2
Transfection of B-lymphoma Cells
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The eGFP-Centrin1 plasmid was obtained from M. Bornens (Institut Curie, Paris, France). The Sec3-HA plasmid was kindly provided by P. Chavrier (Institut Curie, Paris, France). Plasmid expression was achieved by electroporating 2 × 106 B-lymphoma cells with 2 µg of plasmid using the Amaxa Cell Line Nucleofactor Kit R (L-013 program; Lonza). Cells were incubated in CLICK medium for 16–20 h before analysis.
3
Transfection of IIA1.6 B Lymphoma Cells
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+ Expand
The Amaxa Cell Line Nucleo-factor Kit R (L-013 program; Lonza) was used to electroporate 5 × 10 6 IIA1.6 B Lymphoma cells for different transfections. For Centrin-GFP plasmid transfection we used 2 µg of DNA, and cells were cultured for 16 h before functional analysis. In the case of transfection with siRNA we used Silencer® Select (Ambion/Thermofisher) against Nesprin-1(Syne1): s234287 (#1), s234288 (#2); siRNA Sun1: s94911 (#1), s94912 cells (#2), these were incubated for 48 h before analysis. As a control, we used a scrambled siRNA (Qiagen) at 10nM.
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