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Mcherry lamina c 18

Manufactured by Addgene

MCherry-LaminA-C-18 is a fluorescent protein construct that consists of mCherry fused to the C-terminus of human Lamin A protein. Lamin A is a structural protein that is a component of the nuclear lamina. This construct can be used to visualize the localization of Lamin A within the cell nucleus.

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2 protocols using mcherry lamina c 18

1

Vimentin Expression in Inducible Hela Cells

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The Flp-In™ T-REx™ HeLa cells were cultured in Dulbecco’s Modified Eagle’s Medium supplemented with 2 mM GlutaMAX-I, 10% FBS, and 100 U/mL HyClone penicillin-streptomycin at 37 °C and 5% CO2.
The pcDNA5-GFP-Vimetin construct was generated by inserting vimentin cDNA (NCBI Reference Sequence: NM_003380.5) into the pcDNA5-GFP backbone. Plasmid mCherry-LaminA-C-18 (#55068) was ordered at ADDGENE.
The cells were seeded at 1 to 2 × 105 per 6-well, and plasmid transfections were performed using Lipofectamine™ 3,000 transfection reagent. The fluorescent-positive cells were sorted with FACS into a 6-well plate. The positive cells were grown in a selection medium with G418 (200 μg/mL), hygromycin (100 μg/mL), and blasticidin (15 μg/mL). The expression of the vimentin protein was induced with tetracycline at 1 μg/mL for 24 h.
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2

Diverse Fluorescent Protein Constructs

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mcherry, eGFP, eGFP-α-tubulin, eGFP-βactin, and eGFP-Mito were obtained from Clontech. eGFP-VASP [Svitkina et al. 2003 (link)] was a gift from the Borisy lab. eGFP-Utrophin [Burkel et al. 2007 (link)] was a gift from the Bement lab. eGFP-Lifeact [Riedl et al. 2008 (link)] and eGFP-F-tractin [Johnson and Schell 2009 (link)] were gifts from the Baum lab. EMTB-3xGFP [Miller and Bement 2009 (link)] was a gift from William Bement (Addgene plasmid #26741). PM-GFP [Teruel et al. 1999 (link)] was a gift from Tobias Meyer (Addgene plasmid # 21213). EGFP-H2B-6 (Addgene plasmid # 56436), mcherry-LaminA-C-18 (Addgene plasmid # 55068), and mcherry-Vimentin-7 (Addgene plasmid # 55156) [Shaner et al. 2007 (link)] were a gift from Michael Davidson. pcDNA3-HtrA2-EGFP [Martins et al. 2002 (link)] was a gift from L. Miguel Martins (Addgene plasmid # 14121). For simplicity, all text references referring to “GFP” are describing enhanced-GFP constructs.
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