ScreenFectTMA
Contents
1. Screen ectiMA Screen ect A Dilution Buffer 7 NG mg Transfection Reagent for Screen ect A nT O 293 73201 299 73203 Screenfect A 50 00 297 73204 Ref 50ml x 5 200 00 Ref 2 10 C he 3 2013 4 A 540 8605 1 2 TEL 06 6203 1788 103 0023 5 13 TEL 03 3270 8243 092 622 1005 082 285 6381 052 772 0788 029 858 2278 022 222 3072 011 271 0285 Screen Fe ct MA G FP HE K293 o2. URL http www wako chem co jp E mail labchem tec wako chem co jp O 96 G FP 15ng 0120 052 099 0120 052 806 Wako Overseas Offices Wako Chemicals USA Inc Richmond VA Tel 1 804 714 1920 http www wakousa com Sales Offices Los Angeles CA Tel 1 949 679 1700 Boston MA Tel 1 617 354 6772 Wako Chemicals GmbH European Office Tel 49 2131 311 0 http www wako chemicals de 13508 01 Screen ect MA LL wu 0 TT hhh tt ii Li 33 33 1 nas 3 5 6 Screen ect MA tu DAPI A DAPI Relative transfection efficiency GFP Screen ect MA ES GFP GFP ES 60 ScreenFect MA
3. Biomaterials 2012 Nov 33 32 8160 6 2012 GFP Screen ect VA GFP HEK293 HEK293T GFP A DNA Screen ect VA B OKN i te 96 GFP 75ng ScreenfFectIMA G 1 Mix cells Detach and resuspend cells with complexes Combined plating and transfection A lt 4 Screenrect MA A Screenrect MA Screenrect VA ScreenFectMA ScreenFect MA siRNA HEK293 A Mix gently by pipetting Mix gently by pipetting DB E24 Screenrect MA siRNA Screen ect MA 2 O Plasmid DNA Dilution Buffer
4. Screen ectIMA amp DNA amp siRNA a a HEK293 DNA 6 96well 70 98 24 48 h 6 well 20 50 24 48 h EK293 DNA 6 96well 70 98 24 48 h 6welll 70 90 48h Speci c Gene Expression HeL lt 6 24 welll 50 70 24 48 h N 10 20 48 h MC 0 pem Seeee Gone 20 80 24 48 FP 2 52 HepG2 _ 24well 50 70 2h GFP 7 HUVEG DNA amp siRNA 1 Specific Gene 12 well 20 50 48h IRNA 6 well RS MCF 10Al siRNA 6 12well 10 20 48h PLC80 12 well 10 20 48h 1 PLC8022 5 HepG2 50 70 48h ES 6 24 welll 40 60 24 48 h amp Du 10 20 24h GFP HE 6 24 welll 50 70 48h 2013 4 DNA amp siRNA lt lt HEK293 amp 1 rat myoblast
5. VY 10 20 pmol Dilution Buffer Vy 6 75ng 8nL 2min 0 25 1uL of 20uM 40pL 2min 96 pmole LRP6 siRNA 7 2 3 EE 0 i a s 2nM Incubate for 2 5 min at room temperature Incubate for 2 5 min at room temperature 1 Mix gently by pipetting Mix gently by pipetting po a i UH TE dt RE SG Sg Cr K s WE Mix rapidly by pipetting o not vortex Mix rapidly by pipetting o or vortex gt Incubate for 20 30 min at room temperature Incubate for 20 30 min at room temperature SF DNA Complex 20min SF siRNA Complex 20min Add 420uL into SF siRNA Complex Screen ect MA GAPDH Se S Add 80uL into SF DNA Complex J Detached and IS cells by Detached and cells by He La MCF7 A43 1 1 RNA using Trypsin EDTA or a 3 using IS EDTA or Accutase EN DID g Iryp Mix gently by pipetting SI Ih LO SS 4 A Transfer the cells into one well of 96 well plate Transfer the cells into one well of 24 well plate a O i 8 96 GAPDH siRNA DNA transfection well siRNA transfect
6. cells 0 human umbiical vein endothelal cells Screen octTMA 1 Ins 1 murine insulinoma cells human esophageal immortalization cells HeLa HepG2 MDCK Cos 7 ES THP 1 RAW264 7 14 MCF 10a humanmammaryepithelialcels 31 a 15 human liver carcinoma cells DNA siRNA 17 RAW264 7 murine macrophageleukemiacells 34 Primary cells form Liver tissue _ 2013 4 Screen ect WA
7. ion well 9 po 3nM uD cc RS Surface Medium Total volume DNA Screenrect Plat Surface Medium Total volume siRNA Screenrect 5 Q area volume SF DNA complex Dilution Buffer Dilution 0 SSC area volume SF siRNA complex Dilution Buffer Dilution 75ng 0 2 0 4ul 2 3pmol 0 15 0 35ul gt 96 56 wels loam 3cm lt 80Ll 16Ll en 7 300n 1 2ul L 24 wels 2om 350ul 80ul 40 24 wells 420pl 80ul a 1 IP 1 1 _ Reagent u I well 0 04 0 04 0 06 0 09 12 wells 700 140l NB 12 wells 4cm 750ul 150l LM ei 70ul 70pl 75ul siRNA 3nM 1000ng 4 6Ll 20 60pmol 2 5 6Ll Screen ect MA 9 6 wells 10cm 1250ul 240Lul 6 wells 10cm 1260nl 240Ll Eo a 1 NM 120p 120p 120ul 120ul A siRNA Screen ect MA siRNA OKU
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