{"product_id":"cl169","title":"K562-Fluc-EmGFP","description":"\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eParental K562 cells were transduced with \u003c\/span\u003e\u003ca href=\"https:\/\/imanislife.com\/products\/lv-luc2-p2a-puro\/\"\u003e\u003cspan style=\"font-weight: 400;\"\u003eLV-SFFV-Luc2-P2A-EmGFP (Imanis #LV050)\u003c\/span\u003e\u003c\/a\u003e\u003cspan style=\"font-weight: 400;\"\u003e encoding the firefly luciferase (Fluc; Luc2) cDNA under the spleen focus-forming virus (SFFV) promoter and linked to the emerald green fluorescent protein (EmGFP) cDNA via a P2A cleavage peptide. A high Fluc and EmGFP expressing population was generated by selection using a methylcellulose based semi-solid medium.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThis cell line has been tested for mycoplasma contamination and is certified mycoplasma free.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThe parental K562 cell line was licensed and purchased directly from ATCC*.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cb\u003eReplication Competent Lentivirus (RCL) Test \u003c\/b\u003e\u003cspan style=\"font-weight: 400;\"\u003e(including a test report) is available for this cell line at an added cost. \u003c\/span\u003e\u003ca href=\"https:\/\/imanislife.com\/services\/sample-analysis\/\"\u003e\u003cb\u003eContact us\u003c\/b\u003e\u003c\/a\u003e\u003cspan style=\"font-weight: 400;\"\u003e to learn more.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003e*The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cb\u003eReporter Gene Validation\u003c\/b\u003e\u003cspan style=\"font-weight: 400;\"\u003e: \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eMorphology\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003ePhase and fluorescence cell photos taken at 200x magnification\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eLuciferase Activity\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThe indicated number of cells were placed in wells of a 96-well plate. After the addition of 3 mg\/mL d-luciferin, the plate was immediately imaged using an IVIS Spectrum. The total flux (photons\/sec) was plotted as a function of cell number.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eEmGFP Expression\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eK562-Fluc-EmGFP (green) or parental K562 (grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry. \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cb\u003eGrowth Conditions\u003c\/b\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eComplete Growth Medium: Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 15% fetal bovine serum (FBS) and 1% Penicillin\/Streptomycin. \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThese cells should be grown in the indicated medium and subcultured as needed to maintain a density between 5 x 10\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e5\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e and 2 x 10\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e6\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e cells\/mL. The cells can be passaged by dilution in fresh medium, with occasional passaging using centrifugation to limit the amount of debris in cultures.  \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cb\u003eUsage Information\u003c\/b\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThese cells are suitable for \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003ein vitro\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e and \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003ein vivo\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e experimentation. The Fluc transgene facilitates \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003ein vivo\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e noninvasive bioluminescent imaging of implanted cells. EmGFP is not recommended for in-life imaging, but can be used for post-mortem confirmatory assays. Both Fluc and EmGFP facilitate quantitation of cells \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003ein vitro\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e. \u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-weight: 400;\"\u003eThese cells were generated via lentiviral vector transduction. The lentiviral vector used for transduction was a self-inactivating (SIN) vector in which the viral enhancer and promoter have been deleted. Transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization by replication competent viruses and enables regulated expression of the genes from the internal promoters without \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003ecis\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e-acting effects of the LTR\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e1\u003c\/span\u003e\u003cspan style=\"font-weight: 400;\"\u003e. Nevertheless, all work with these cells should be performed under biosafety-level 2 (BSL2) conditions by trained personnel. Institutional requirements may permit handling of these cells under BSL1 conditions if certain criteria are met.\u003c\/span\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli style=\"font-weight: 400;\"\u003e\n\u003cspan style=\"font-weight: 400;\"\u003eMiyoshi, H \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003eet al. \u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e(1998). Development of a self-inactivating lentivirus vector. \u003c\/span\u003e\u003ci\u003e\u003cspan style=\"font-weight: 400;\"\u003eJournal of Virology\u003c\/span\u003e\u003c\/i\u003e\u003cspan style=\"font-weight: 400;\"\u003e 72: 8150-8157. \u003c\/span\u003e\n\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003cbr\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003col data-mce-fragment=\"1\"\u003e\u003c\/ol\u003e","brand":"Imanis Life Sciences","offers":[{"title":"Frozen \/ Standard","offer_id":42617407439029,"sku":"CL169-STAN","price":3000.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0683\/0295\/products\/k562-fluc-emgfpmorphology.png?v=1674767543","url":"https:\/\/imanis-life-sciences-4.myshopify.com\/products\/cl169","provider":"Imanis Life Sciences","version":"1.0","type":"link"}