Lipofectamine 3000 pdf

Lipofectamine® 3000 Reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells and especially designed for hard to transfect cells Important Guidelines ∤ Make DNA-Lipofectamine® 3000 complexes in serum-free medium such as Opti-MEM® Reduced Serum Medium and add directly to cells in culture ... dance with the Lipofectamine® 3000 transfection reagent instruction. The miR-26a group only transfected miR-26a mimics, the miR-NC group only transfected miR-26a negative control mim-ic, the co-transfection group transfected miR-26a mimics and pcDNA3.1-SMAD1 plasmid, and the blank control group was not transfected. The packaging mix, using Lipofectamine 3000 reagent (Invitrogen) to generate retroviral particles. After 48– 72 h of transfection, the medium containing the retrovirus was harvested and transduced into the target cells in the presence of polybrene (8 μg/mL). Transfected cells were selected by culturing for 2

transfection was operated using lipofectamine 3000 (Invitrogen, USA) according to standard protocol. A total of 20μMsi−/over-expressing-RNA and empty con-trol plasmids were respectively transfected using 3.75μl lipofectamine 3000 diluted in Opti-MEM and incubated at room temperature for 15min. After being cultured 3000. 4000. OC442107a - 10/2017. 1000. 2000. 3000. a. b c.3000. 4000. OC442107a - 10/2017. 1000. 2000. 3000. a. b c.CaSKi cells were performed using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) by si-CALML3-AS1 or si-NC. Real Time-PCR Total RNAs were extracted employing TRIzol reagents (Invitrogen, Carlsbad, CA, USA). cDNA was reversely transcribed from collected RNAs (2 mg) by the use of M-MLV reverse transcrip-tase (Invitrogen, Carlsbad, CA, USA).

Lipofectamine 2000 and Lipofectamine 3000, performed poorly in basal media, yielding less than 25% of the fluorescence intensity as that of the XtremeGENE HP. The differences in transfection effi-ciencies among the reagents tested were further evident under epi-fluorescent microscopy. Figure 1B shows epi-fluorescence images of Download PDF Download PDF. Original Version Original Version Updated Version Updated Version. The transfection procedure is performed using Lipofectamine3000 reagent (Invitrogen) according to...combined with Lipofectamine 3000 transfection reagent, we can achieve about 90% Cx43 knockdown efficacy in HUVECs. Therefore, our results suggest that a much higher transfection and viral transduction efficiency can be at-tained by using Lipofectamine 3000 transfection reagent.

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Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. A low toxicity lipid dose (0.75µL for 24-well plates) is suggested for applications requiring minimal disruption of the cells. Lipofectamine 3000 Transfection Reagent 0.1 mL 15282465 Lipofectamine RNAiMAX Transfection Reagent 0.1 mL 12333563 Lipofectamine MessengerMAX Transfection Reagent 0.3 mL 15397974 ExpiFectamine 293 Transfection Kit 1 x 1 L culture 13385544 Lipofectamine 2000 Transfection Reagent 0.3 mL 12313563 Invivofectamine 3.0 Reagent 10 reactions 15331018 Nov 30, 2020 · 142 using Lipofectamine 3000 transfection reagent (ThermoFisher Scientific) with 800 ng of DNA 143 (400 ng WT and mutants, respectively, for co-expression transfection). The medium was changed 144 24 h after transfection. Cells were harvested 72 h after transfection and solubilized in radio-

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Detailed instructions on reagent preparation and assay protocol for an MTT assay to measure cell proliferation or cell cytoxicity.

Cells were transfected using Lipofectamine 3000 transfection reagent (Invitrogen) according to the manufacturer’s protocol. VeCtoRs pCMV6-PRMT1, pCMV6-DDAH1, pCMV6-DDAH2 vectors were from Origene. AAV8.TBG. PI.Null, AAV8.TBG.PI.Cre were from Vector BioLabs (Malvern, PA) and were used at 1-2 × 1011 genome copies (gc)/mouse. Human speCimens Кофемашина. EAM 3000 esam 3000 EAM 3100 esam 3100 EAM 3200 esam 3200. àÌÒÚÛ͈ËË ÔÓ ˝ÍÒÔÎÛ‡Ú‡ˆËË. S t k lu a.

Title: 11814443001a.fm Author: knepflem Created Date: 3/15/2006 4:21:24 PM Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). INS-1E cells or pancreatic islets were treated with cytokine cocktail (CKS), consisting of TNF-α (50 ng/ml), IL-1β (100 ng/ml) and IFN-γ (100 ng/ml), or PBS containing 0.1% wt/vol. fatty-acid-free BSA as vehicle. For details of islet isolation, see ESM Methods.

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  1. fectamine 3000/FAM-siRNA complexes. At 250 nM, the transfection did not change notably. Therefore, we decided to use 200 nM of siRNA as the optimal dose for HNT transfection in T24 cells. The use of HNTs induced similarly low toxicity to that of Lipofectamine 3000 during siRNA transfection. Fig. 1. Characterization of the siRNA-loaded HNTs.
  2. CHO-K1 cells were transfected with EGFP DNA and Lipofectamine. Control. 500 3000 bp 400 Impurity level : > 50%.
  3. Invitrogen™ Lipofectamine™ 3000 Transfection Reagent is a highly efficient, cost-effective tool for Advantages of lentiviral production using Lipofectamine 3000 reagent: • Superior performance...
  4. Download PDF. Research. Open Access. Diluted DNA was added dropwise to the diluted Lipofectamine™ 3000 reagent at a 1:1 ratio and incubated for 10 min at room temperature.
  5. Mar 21, 2018 · rat cardiac myocytes with Lipofectamine 3000 (Thermo Fisher, USA). The Gata4-small interfering RNAs sequence was 5 0-GGCCUCUAUCACAAGAUGA-3 . Transfection of Adeno-Associated Virus Adeno-associated virus subtype 9 containing Gata4-shRNA (AAV9-Gata4-shRNA) and negative-control vectors (AAV9-NC) were purchased from Genepharma (Shanghai, China). The
  6. Table 1: Lower volume of jetOPTIMUS ® is required compared to Lipofectamine ® 3000 in 24-well plate.. Excellent cell viability and morphology. Reaching high transfection efficiencies is often at the expense of cellular integrity, with adverse effects on cell cycle, metabolism and signaling pathways.
  7. transfection reagent Lipofectamine® 3000. We describe this new technology and how this simple versatile reagent delivers efficient, reporducible transfection of both DNA and RNA ino a wide range of hard-to-transfect, biologically relevant cell models. In the present study, we demonstrated that Lipofectamine® 3000 can be used to
  8. NHJB-3000-P (01/01/2017) Page 2 of 2 8. The Person is represented by counsel (if known to the Petitioner): Name Mailing Address 9. The Petitioner proposes that the individual listed below, who is not an employee of the Petitioner, be appointed by the Court as the Person’s Special Medicaid Representative: Name Mailing Address
  9. NHJB-3000-P (01/01/2017) Page 2 of 2 8. The Person is represented by counsel (if known to the Petitioner): Name Mailing Address 9. The Petitioner proposes that the individual listed below, who is not an employee of the Petitioner, be appointed by the Court as the Person’s Special Medicaid Representative: Name Mailing Address
  10. Lipofectamine™ 3000 reagent was purchased from Invitrogen Corporation (Carlsbad, CA, USA). Antibodies against phospho-ERK and ERK were obtained from Cell Signaling Technology (Beverly...
  11. PROTOCOL The following is a general protocol for use of DharmaFECT™ transfection reagents to deliver siRNA into cultured mammalian cells. The examples
  12. CaSKi cells were performed using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) by si-CALML3-AS1 or si-NC. Real Time-PCR Total RNAs were extracted employing TRIzol reagents (Invitrogen, Carlsbad, CA, USA). cDNA was reversely transcribed from collected RNAs (2 mg) by the use of M-MLV reverse transcrip-tase (Invitrogen, Carlsbad, CA, USA).
  13. Lipofectamine 2000 (Invitrogen). 2.4. Subcellular fractionation Cells were collected and resuspended in hypotonic buffer (10 mM KCl, 210 mM sucrose, 70 mM mannitol, 1 mM EDTA, 1 mM EGTA, 1.5 mM MgCl 2, 10 mM HEPES; pH adjusted to 7.4 with KOH). After gentle homogenization with a Dounce homogenizer
  14. pool DNA, psPAX2 and pMD2.G using lipofectamine 3000. Identification of drug-resistant mutations from the random mutation pool BaF3 cells (1 × 107 cells) (Huanget al., 2016) were cultured in RPMI-1640/10% FBS/1 ng·mL 1 mouse IL-3 and infected with the lentiviruses prepared above. Infected cells were selected with puromycin (1 μg·mL 1 ...
  15. Calculus 3000 Solved Problems in Calculus 3000 Solved Problems in Ca ... 3000-WORD ENGLISH-FRENCH VOCABULARY This book is a learning tool for mastering basic French ...
  16. For most cell lines, Lipofectamine LTX (Life Technologies) works well with the manufacturer's suggested protocol. Centrifuge the lysate at 3000 rpm at 4 C for 5 min to pellet nuclei.
  17. Lipofectamine® 3000 is a versatile reagent that can also be used to deliver siRNA using the same transfection protocol. Simply substitute siRNA for DNA. For this experiment, knockdown of endogenous luciferase was achieved in three engineered luciferase cell lines using Lipofectamine ® 3000, Lipofectamine ® 2000, and
  18. A short summary of this paper 34 Full PDF related to this paper These linear polymers exhibited a strong DNA PLL and Lipofectamine 2000 in both COS-7 cell...
  19. Abstract. Atopic dermatitis (AD) is a relapsing inflammatory skin disease with a complicated pathogenesis. This study aimed to investigate whether miR-375-3p co
  20. Jul 26, 2016 · There are several ways in which to introduce Cas9-guide RNA complexes into cells. Here we present a method for the transfection of Cas9 RNP’s into HEK293 FT cells using Thermo Fisher Lipofectamine ® RNAiMAX. This is a ‘reverse transfection’ method that uses a final concentration of 10 nM RNP per transfection in a 96-well culture plate.
  21. poGene.pdf. 540.50 KB; Cite. 9 Recommendations. All Answers (12) 6th Jul, 2015. ... This is the first time I will have used Lipofectamine 3000, previously I only used Lipofectamine 2000, and I am ...
  22. centrifugation for 10 minat 3000 rpm, the chloroform layer was collected into a new screw cap tube through filter paper. The amount of phospholipids was quantitated by phosphorus assay (37), and dimyristoleoyl PC (Avanti Polar Lipids, Birmingham, AL, USA) was added as an internal standard. The liquid
  23. Lipofectamine 3000这个成分,在质粒转染中起主要作用,这个东西加少了,转染效率下降,加多了,对细胞的毒性变强。所以,我们想要找到一个平衡点,让转染效率比较高的同时,细胞毒性不太大。因此,在正式的批量细胞转染实验进行之前,需要安排一套预实验 ...
  24. pool DNA, psPAX2 and pMD2.G using lipofectamine 3000. Identification of drug-resistant mutations from the random mutation pool BaF3 cells (1 × 107 cells) (Huanget al., 2016) were cultured in RPMI-1640/10% FBS/1 ng·mL 1 mouse IL-3 and infected with the lentiviruses prepared above. Infected cells were selected with puromycin (1 μg·mL 1 ...
  25. IMLF can be transfected using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA) but transfection efficiency is higher using recombinant adenovirus (described later), which gives very good expression in a short-time relative to other methods. 1.
  26. were transfected to cells with Lipofectamine 3000 (Life Technologies,Waltham, MA). Cells were harvested for 48 hours after transfection before mRNA extraction or ChIP assay. All siRNAs were synthesized in GenePharma (Shanghai, China); the target sequences were listed in Supplemental Table 4. Coimmunoprecipitation.
  27. Lipofectamine 3000 reagent** siRNA Lipofectamine 3000 reagent** 96-well 0.2 100 μL 2 x 5 μL 50 ng 0.1 µL 0.15 µL 3 pmol 0.3 µL 48-well 0.5 250 μL 2 x 12.5 μL 0.125 µg 0.25 µL 0.375 µL 7.5 pmol 0.75 µL 24-well 1 500 μL 2 x 25 μL 0.25 µg 0.5 µL 0.75 µL 15 pmol 1.5 µL 12-well 2 1 mL 2 x 50 μL 0.5 µg 1 µL 1.5 µL 30 pmol 3 µL

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  1. GenePharma (Shanghai, China). Lipofectamine 3000 (Invitrogen) was used to transfect cells with miRNAs and HMGA2 for subsequent experiments according to the manufacturer’s instructions. The sequences of the shRNA clone and the agomir and antagomir are listed in Additional file 1: Table S1. RNA extraction and quantitative RT-PCR
  2. 3000/4000. Allison Transmission VOCATIONAL MODELS Operator’s Manual 2005 FEBRUARY Rev. 1 2005 SEPTEMBER OM3349EN Motorhome Series (MH) Transmissions
  3. Lipofectamine RNAiMAX Transfection Reagent provides the highest transfection efficiencies on the widest variety of cell types for siRNA mediated gene knockdown experiments Lipofectamine RNAiMAX is a proprietary RNAi specific cationic lipid formulation designed specifically for the delivery of siRNA and miRNA into all cell types With ...
  4. 2000 and Lipofectamine 3000 were used to transfect U2OS and HepG2 cells in a 12-well format. Effi ciency and OFP expression were analyzed 72 hours posttransfection and (A) U2OS and (B) HepG2 cells showed 4-fold and 80-fold improvement with Lipofectamine ®
  5. Scaling Up or Down Lipofectamine® 3000 Reagent Transfections Use the following table to scale the volumes for your transfection experiment. The most common sizes factor are listed below. 1 After determining the optimum reagent amount, use the multiplication factor to determine the reagent amount needed for your new plate format.
  6. TRANSFECTION PROTOCOL Lipofectamine 3000 Transfection Reagent MCF10A Breast cancer cells Complete growth medium Component Cat. No. Gibco™ Ham’s F-12 Nutrient Mix 11765054 5% Horse Serum – 1 mM Gibco™ Sodium Pyruvate 11360070 25 mM Gibco™ HEPES 15630106 Proper culture techniques and procedures are an essential
  7. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. A low toxicity lipid dose (0.75µL for 24-well plates) is suggested for applications requiring minimal disruption of the cells.
  8. Mar 12, 2018 · Signal Transduction Definition. Signal transduction is the process of transferring a signal throughout an organism, especially across or through a cell.Signal transduction relies on proteins known as receptors, which wait for a chemical, physical, or electrical signal.
  9. A B DNA: 1.0 µg Lipofectamine 3000: 1.5 µL SSEA+/GFP+: 42% GFP MFI 247344 DNA: 1.3 µg Lipofectamine 3000: 1.5 µL SSEA+/GFP+: 69% GFP MFI 456741 ® ® 図6. 幹細胞へのトランスフェクション Lipofectamine® 3000を用いて、H9 ESC( A)またはiPSC(B)にトランスフェクションを行いました。
  10. PROVOST’&’WALLERT’RESEARCH! Investigating the Biochemistry & Cellular Physiology of NHE1 EST. 1998 Stable Cell Line Production! UpdatedSpring2012KA!
  11. Mitochondrial unfolding protein response (UPR<sup>mt</sup>) effectively resists the pathological cardiac hypertrophy and improves the mitochondrial function. However, the specific activation mechanism and drugs that can effectively activate UPR<sup>mt</sup> in the cardiac muscle are yet to be elucidated. The aim of this study was to determine the regulation role of UPR<sup>mt</sup> on ...
  12. CA, USA). The streptavidin-conjugated 605QDs, Lipofectamine™ 3000 transfection reagent and Opti-MEM® I reduced-serum medium were purchased from Thermo Fisher Scientific (Massachusetts, USA). The miRNeasy RNA isolation kit was obtained from Qiagen (Valencia, CA, USA).
  13. Oct 06, 2017 · 2 × 10 7 HeLa cells (2 × 15 cm plates) were subjected to 72 hr knockdown using Lipofectamine RNAiMAX according to the manufacturer's protocol. After 72 hours, cells were washed with PBS and collected with a Cell Lifter (Corning). 10% was used for analysis of total RNA, and the remainder was separated into cytoplasmic and nuclear components.
  14. Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics with His-tag (Vigene Biosciences, Inc.) and the empty vector were transfected into H1299 cells using Lipofectamine® 3000.
  15. Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. Figure 2. The transfection protocol for Lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines.
  16. Transfer viral supernatnat to Amicon Filter and spin filter in tabletop centrifuge at 3000 rpm for 10-20 min at 4C. Concentrated virus can be aliquoted and stored at -80C. Thaw an aliquot on ice before use; do not refreeze. Concentration by ultracentrifugation: 1. Transfer viral supernatant to 33ml Beckman ultracentrifugation tubes and spin for 2
  17. RNA oligonucleotides were transfected by using Lipofectamine 2000 (Invitrogen) and the medium was replaced 6 hours after transfection. Cy3-labeled miRNAs were transfected into macro-phages with using Lipofectamine 3000 (Invitrogen). Macro-phages containing Cy3 miRNA were cocultured with SW48 cells
  18. Add Lipofectamine 2000 to Opti-MEM in. another 1.5 ml tube (see table for amounts) and gently 5. Remove Optimum medium from cell dish 6. Add DNA/Lipofectamine 2000 diluted mixture on cell with...
  19. lipofectamine 3000 (Thermo Fisher Scientific). Twenty-four hours after transfection, cells were fixed in 4% para-formaldehyde, and immunofluorescence was performed as described previously9 using M2 anti-FLAG antibody (Sigma, 1:250) and sodium potassium ATPase antibody (Abcam 76020,1:100). Nuclear staining was performed using DAPI (BioLegend ...
  20. Jun 15, 2020 · Lipofectamine™ 3000 Transfection Reagent (Invitrogen)in 293T cell s. The collected supernatants were collected and purified by protein G-sepharose (Thermo Fisher). The . available under aCC-BY-NC-ND 4.0 International license.
  21. Transfection Methods www.bio-rad.com/transfection Factors Affecting Transfection 5 Host Cell Cell health Cell culture Genetic Material DNA quality and quantity

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