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LipoJet™ (Ver. II) DNA In Vitro Transfection Reagen
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ϸ°ûÀàÐÍ | DNAתȾЧÂÊ | siRNAתȾЧÂÊ |
293, 293T | 85% | - |
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A efficiency comparison of LipoJet™ reagent vs. brand name products to transfect Hela, Cos-7, NIH-3T3, CHO £ánd HEK293 (left panel) £ánd Cos-7 (right panel). pEGFP-N3 cDNA (0.5 µg/well of 24-well plate) was transfected into different mammalian cells per the standard transfection protocols in presence of serum (10% FBS) £ánd antibiotics as recommended by manufacturers. The transfection efficiency were analyzed via FACS 48 hours post transfection.
A toxicity comparison of LipoJet™ reagent vs. brand name products to transfect Hela cells. pEGFP-N3 cDNA (1.0 µg/well of 24-well plate) was transfected to Hela cells per the standard transfection protocols in presence of serum (10% FBS) £ánd antibiotics as recommended by manufacturers. The MTT assay (right panel) £ánd phase contrast imaging (left panel) were used to analyze the cell viability 48 hours post transfection.
A comparison of LipoJet™ reagent vs. Lipofectamine 2000 (L2K) £ánd PolyJet™ transfection reagent on HEK293T cell. pEGFP-N3 cDNA (0.125 µg/well, 0.25 µg/well £ánd 0.5 µg/per well of 24-well plate) was transfected into 293T cells using the standard transfection protocols in presence of serum (10% FBS) with LipoJet™ (upper panel at LipoJet™/DNA (µl/µg) ratio=2), L2K (middle panel at L2K/DNA (µl/µg) ratio=3) £ánd PolyJet™ (lower panel at at PolyJet™/DNA (µl/µg) ratio=3) respectively. The cells were visualized by Nikon Eclipse Fluorescence microscope 48 hours post transfection.
A comparison of LipoJet™ reagent vs. Lipofectamine 2000 (L2K) £ánd PolyJet™ transfection reagent on Hela cell. pEGFP-N3 cDNA (0.125 µg/well, 0.25 µg/well £ánd 0.5 µg/per well of 24-well plate) was transfected into Hela cells using the standard transfection protocols in presence of serum (10% FBS) with LipoJet™ (upper panel at LipoJet™/DNA (µl/µg) ratio=2), L2K (middle panel at L2K/DNA (µl/µg) ratio=3) £ánd PolyJet™ (lower panel at at PolyJet™/DNA (µl/µg) ratio=3) respectively. The cells were visualized by Nikon Eclipse Fluorescence microscope 48 hours post transfection.
LipoJet™ÊÔ¼Á½éµ¼µÄDNA/siRNA¹²×ªÈ¾±íÏÖ³öÉ«µÄ»ùÒò³ÁĬЧӦ¡£·Ö±ðÓÉLipoJet™ÊÔ¼ÁתȾGFPµÄcDNA (×óͼ, 0.25µg/¿×£¬24¿×°å)×÷Ϊ¶ÔÕպ͹²×ªÈ¾GFPµÄcDNA (0.25µg/¿×£¬24¿×°å)/GFPµÄsiRNA (ÖÕŨ¶È10 nM£¬ÓÒͼ)ÖÁHEK293ϸ°û (ÉÏͼ), Helaϸ°û(ÖÐͼ)ºÍSaoS-2ϸ°û(ÏÂͼ)¡£×ªÈ¾24Сʱºó£¬Ê¹ÓÃÄ῵EclipseÓ«¹âÏÔ΢¾µ¼ì²éGFPÓ«¹â¡£½á¹ûÏÔʾ£¬GFPµÄcDNAºÍGFPµÄsiRNA¹²×ªÈ¾µ¼ÖÂGFPµÄ±í´ï±»ÏÔÖøÒÖÖÆ¡£
Exceptional DNA/siRNA co-transfection efficiency on HUVEC. Co-transfection of mCherry cDNA (0.10 µg per well of 24-well plate) £ánd FITC conjugated siRNA (final 30 nM per well of 24-well plate) to HUVEC with LipoJet™ reagent gave rise to 60% mCherry+ (phase contrast overlapped with mCherry imaging, left panel) £ánd nearly 100% FITC-siRNA+ (phase contrast overlapped with FITC imaging, right panel) HUVEC 24 hours after transfection. The pictures were given from Dr. Pan Kong of USC as courtesy.
Excellent silencing of endogenously expressed KIF11 (also known as EG5) in HEK293 (upper panel) £ánd Hela (lower panel) cells with LipoJet™ reagent at 10 nM EG5 siRNA. KIF11 (also known as EG5) encodes a motor protein that belongs to the kinesin-like protein family involved in chromosome positioning £ánd bipolar spindle formation during cell mitosis. A reduction in KIF11 levels causes mitotic arrest. LipoJet™ reagent effectively delivers EG5 siRNA (final 10 nM) to HEK293 £ánd Hela cells, leading to more than 80% of "round-up" phenotype of HEK293 £ánd Hela cells 24h post transfection over negative control (final 10 nM with sham EG5 siRNA). The phenotype of "rounded-up" HEK293 £ánd Hela cells were visualized 24h post transfection with a Nikon microscope.