Transfection Reagents That Really Work

Transcription

1 Transfection Reagents That Really Work non-toxic, biodegradable, highly efficient, serum-compatible Available for DNA,, protein, stem cells

2 P L A S M I D T R A N S F E C T I O N R E A G E N T S Transfection Reagent Simple, serum-compatible protocol High transfection efficiency Very low cytotoxicity Broad range of cells tested (see back of brochure) Simple, Serum-Compatible Protocol For ease of use and happier cells, plasmid transfections can be carried out entirely in the presence of serum. The transfection protocol requires few steps and minimal optimization and there s no need to mess around with ratios! The reagent kit includes everything you need no Opti-MEM is required. Plasmid DNA solution Reaction buffer + plasmid DNA Combine & vortex Incubate, then apply to subconfluent cells Incubate 4 hr to overnight, then remove & replace with fresh media Assay for gene expression 48 hr posttransfection Polymer solution Reaction buffer + polymer High Transfection Efficiency & Cell Viability If you want to increase transfection efficiency, it makes sense to increase the amount of plasmid DNA and transfection reagent that you add to your cells. Unfortunately, most transfection reagents damage cells during transfection, which makes this impossible. is different. is biodegradable and nontoxic, so you can transfect more DNA without killing your cells. Excellent Results in Many Cell Lines transfects most commonly-used cell lines with very high efficiency (see back cover). It even works well in Jurkat cells, which are notoriously difficult to transfect: transfects Jurkat cells with 28% efficiency, while X transfects them with just.65% efficiency. Convenient Multipack Format Viable transfected cells (% of total) ng 5 ng 1. μg 2.5 μg 5. μg 7.5 μg 1 μg Amount of DNA transfected The Transfection Reagent is packaged in convenient, separable kits, so you can share with your less-than-careful colleagues without fear. Simply give them part of the kit, and hide yours!

3 P L A S M I D T R A N S F E C T I O N R E A G E N T S Adult Stem Cell Transfection Reagent Higher efficiency than the leading competitors Up to 67% transfection efficiency in human mesenchymal stem cells Up to 99% transfection efficiency in human adipose-derived stem cells Does not affect differentiation Adult stem cells have great potential therapeutic value because of their multipotency. We optimized the Adult Stem Cell Transfection Reagent to maximize transfection efficiency and retain low cytotoxicity in adult stem cells, without affecting differentiation. Adult Stem Cell Transfection Reagent Competitor L Competitor F Human adipose-derived stem cells were transfected with a plasmid encoding AcGFP1 using three different transfection reagents (see right). The Adult Stem Cell Transfection Reagent demonstrated far higher transfection efficiency than Competitors L or F. Mouse Embryonic Stem Cell (mesc) Transfection Reagent Better efficiency than the leading competitors Superior gene expression in mes cells Simple, serum-compatible protocol In a head-to-head comparison with three other reagents, mesc provided the best transfection efficiency in multiple mes cell lines, including ES-E14TG2a and ES-D3. Like the original, the mesc Transfection Reagent has a low cytotoxicity profile and excellent transfection efficiency. % Transfection efficiency E14TG2A mes cell line mesc X Product F D3 CalPhos Calcium Phosphate Transfection Reagent High efficiency, dependable results Optimized ph and salt concentration 7 transfections (6-well plate format) Calcium phosphate transfection is a common method for delivering plasmid DNA into mammalian cells. Due to its highly consistent ph and salt concentration, the CalPhos Mammalian Transfection Kit gives reliable transfection efficiencies that you can t always achieve with home brew transfection reagents.

4 s i R N A T R A N S F E C T I O N R E A G E N T S Transfection Reagent Better knockdown than the leading competitor No optimization required Nontoxic won t kill your cells Effective Gene Silencing Many transfection reagents were designed for plasmid delivery and then sold for transfection. The Transfection Reagent was specifically created and optimized for delivery so it provides better results than all-purpose reagents like. PP1B expression in HeLa cells Lamin A/C expression in HeLa cells MAPK1 expression in HeLa cells Relative mrna level Relative mrna level Relative mrna level PP1B control Lamin A/C control MAPK1 control High Viability The Transfection Reagent maintains a very low cytotoxicity profile while providing high rates of gene silencing. Don t hesitate to transfect your cells will survive! (We routinely measure 1% viability for HeLa cells.) Relative level of PP1B mrna T HT18 Cell line HeLa Cell line Knockdown efficiency Relative cell viability 293T 6% 97% HT18 75% 87% HeLa 93% 12% Anti-PP1B control No Optimization Required Many transfection reagents require you to pre-test different ratios of oligo to reagent. But not the Transfection Reagent we ve pre-determined the best ratio for any size of plate or dish, so you can skip the preliminaries and get started with your experiment!

5 P R O T E I N T R A N S F E C T I O N R E A G E N T S Protein Transfection Reagent Faster than DNA transfection, because it bypasses transcription & translation Ideal for studying transient effects of proteins Avoids potentially harmful, random DNA integration into target cells genomes High Transfection Efficiency The Protein Transfection Reagent is more efficient than Product C across a broad range of mammalian cell types, including a number of rodent and human cell lines, hard-to-transfect human suspension cells, and mouse embryonic stem cells. 1 8 Protein Product C Simple Protocol The Protein Transfection Reagent is a modified peptide that interacts with a protein cargo and transports it across the cell membrane barrier. Just incubate your protein of interest with in the supplied buffer, apply the mixture to your cells, and assay for protein activity. You ll get results faster, because the Protein reagent bypasses the requirement for transcription and translation. Untransfected HeLa Cells Product C Positive cells (%) NIH3T3 HEK293 HT18 HeLa CHO-K1 HL-6 Jurkat E14TG2A mes Why Use the Protein Transfection Reagent? It s more effective! Shortfalls of other reagents Cytotoxicity Low efficiency You must transfect cells at low density, so you don t get enough material for analysis. The Protein solution Unlike other lipid-based reagents, Protein has low cytotoxicity. Compared to other peptide-based reagents, Protein delivers more protein, to more cells. Protein can transfect cells growing at a high density, ensuring sufficient material for downstream analyses.

6 T R A N S F E C T I O N R E A G E N T S Can I Transfect my Cells with? Yes! The plasmid transfection reagent has been used (by customers) on all of the following types of cells: Fibroblasts BHK-21, BJ fibroblasts, chick fibroblasts, COS-7, HT18, mouse embryonic fibroblasts (MEF), NIH3T3, Rat2, swiss mouse embryonic fibroblasts Epithelial Cells 16HBE, A431, As4.1, B16-F1, BEAS-2B, BSC1, Caco-2, EpH4, HaCaT, HC11, HEK-293, HEK-293T, hose, HT-18, HT-29, human lens epithelial cells, MCF7, MDCK, T-47D, WRL 68 Endothelial Cells Aortic endothelia (human), HUVEC, myoendothelia (human) Bone Cells MC3T3, U2-OS Colon Cells Caco-2, HT-29 Immune Cells Jurkat, DC2.4, K-562, RAW 264.7, RBL-2H3, SupT1 Kidney Cells 293FT, As4.1, BHK-21, B-S-C1, COS-7, HEK-293, HEK-293T, renal proximal tubules (mouse) Lung Cells 16HBE, BEAS-2B Muscle Cells C2C12, aortic smooth muscle (human), cardiomyocytes (primary rat), myoblasts (primary human), smooth muscle cells (primary human) Neural-Based SH-SY5Y, HEI-193 (human Schwann) Stem Cells hadsc (adipose), hmsc (mesenchymal), mesenchymal progenitor, P19 Reproductive Organs CHO, EpH4, HC11, HeLa, hose, LbT2, MCF7, PC3, T-47D Ordering Information Cat. # Product Package Size Plasmid Transfection Reagents Transfection Reagent 1 rxns rxns Adult Stem Cell Transfection Reagent 1 rxns mesc Transfection Reagent 1 rxns rxns CalPhos Mammalian Transfection Kit 7 rxns Transfection Reagent Transfection Reagent 1, rxns Protein Transfection Reagent Protein Transfection Reagent 3 rxns rxns Takara Bio Europe SAS Clontech Laboratories orders@clontech-europe.com tech@clontech-europe.com Europe : +33.() Austria : Germany : Switzerland : United Kingdom : Notice to Purchaser Your use of these products and technologies is subject to compliance with any applicable licensing requirements described on the product s web page at It is your responsibility to review, understand and adhere to any restrictions imposed by such statements. For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resale. Clontech, the Clontech logo, are trademarks of Clontech Laboratories, Inc. All other marks are the property of their respective owners. Certain trademarks may not be registered in all jurisdictions. 212 Clontech Laboratories, Inc. This brochure is printed on 6% recycled paper. ZZXBR633463EU