Colorful choices for your bright ideas

Living Colors Fluorescent Proteins Reporters and Antibodies for Every Application Colorful choices for your bright ideas

Living Colors Fluorescent Proteins Full color spectrum cyan to far red Ideal for real-time and multicolor imaging Optimal fluorescence and photostability Extensive array of vector formats Suitable for a range of applications Specific fluorescent protein antibodies Easily detected and tolerated in live cells Clontech s Full Collection of Living Colors Fluorescent Proteins Fluorescent Protein Color Excitation Maximum (nm) Emission Maximum (nm) Relative Quantum Yield a Extinction Coefficient (M 1cm 1) b Brightness c Recommended Filter Set(s) d mplum far red 590 649 0.10 41,000 4,100 C.T. 41043 E2-Crimson far red 611 646 0.23 126,000 28,980 C.T. 41019 mraspberry far red 598 625 0.15 86,000 12,900 C.T. 41043 HcRed1 far red 588 618 0.03 20,000 600 C.T. 41043; Omega XF102-2 mcherry red 587 610 0.22 72,000 15,840 C.T. 49008; Omega XF102-2 mstrawberry red 574 596 0.29 90,000 26,100 C.T. 42004 & 42005 AsRed2 red 576 592 0.21 61,000 12,810 C.T. 42004 & 42005 DsRed-Monomer red 557 592 0.14 27,300 3,822 C.T. 41002c & 42005 DsRed2 red 563 582 0.55 43,800 24,090 C.T. 41002c & 42005 tdtomato red 554 581 0.69 138,000 95,220 C.T. 41002c & 42005 DsRed-Express red 554 586 0.44 33,800 14,872 C.T. 41002c & 42005 DsRed-Express2 red 554 591 0.42 35,600 15,000 C.T. 41002c & 42005 morange orange 548 563 0.69 71,000 48,990 C.T. 49010 morange2 orange 549 565 0.60 58,000 34,800 C.T. 49010 mbanana yellow 540 553 0.70 6,000 4,200 C.T. 42003 & 41028 ZsYellow1 yellow 529 539 0.65 20,000 13,000 C.T. 42003 & 41028 ZsGreen1 green 493 505 0.91 43,000 39,130 C.T. 42002 & 41017 AcGFP1 green 475 505 0.82 32,500 26,650 C.T. 42002 & 41017 Dendra2 (converted) Timer (converted) PAmCherry (activated) green (red) green (red) (red) 490 (553) 483 (558) 404 (564) 507 (573) 500 (583) (595) 0.50 (0.55) ND e (ND) (0.46) 45,000 (35,000) ND e (ND) 6,500 (18,000) 22,500 (19,250) ND e (ND) (8,280) C.T. 42002 & 41017 (C.T. 41002c & 42005) C.T. 42002 & 41017 (C.T. 41002c & 42005) C.T. 49008 AmCyan1 blue 458 489 0.75 39,000 29,250 C.T. 42001 & 41014 EYFP g yellow 512 529 0.54 45,000 24,300 EGFP g green 484 510 0.70 23,000 16,100 ECFP g blue 439 476 0.15 20,000 3,000 a The Relative Quantum Yield of each fluorescent protein (Q FP ) was determined using the following equation: Q FP = (Q fluo * F FP * OD fluo ) / (F fluo * OD FP ), where F is the fluorescence and OD is the optical density of either the fluorescent protein (FP) or the reference fluorophore (fluo). b The Molar Extinction Coefficient (e) for each protein was determined as follows: A = ecl (Beer-Lambert law) or e = A/c, where A = absorbance, c = concentration (determined by the Bradford Protein Assay), l = path length (1 cm). Each molar extinction coefficient was determined at the absorption maxima of its respective protein. c Brightness = (Quantum Yield)(Extinction Coefficient) d C.T. = Chroma Technologies e ND = not determined f NA = not applicable g For reference only; not sold by Clontech

Applications of Living Colors Fluorescent Proteins Clontech s fluorescent proteins are available in plasmid, lentiviral, and retroviral vector formats for a range of key applications. Visit www.clontech.com for a complete list of our vectors. Fusions FRET Cellular & subcellular labeling Gene expression Kinetic studies Cell capture Cell cycle reporters Proteasome activity analysis Promoter reporters Timed gene expression In vivo imaging Stem cell applications Selected Applications of Living Colors Fluorescent Proteins Application Recommended Products Comments Protein localization Fusion fluorescent proteins from far red to cyan are available in a variety of vector formats for both N- and C-terminal fusions. AcGFP1, DsRed-Monomer, and mcherry proteins are ideal for fusions, since they are monomeric and so are least disruptive to the function of the protein of interest. Bacterial expression Subcellular labeling In vivo imaging Fluorescence resonance energy transfer (FRET) Protein dynamics & half-life We offer a variety of basic fluorescent protein vectors for bacterial expression. AcGFP1 DsRed-Monomer DsRed2 mcherry HcRed1 mplum E2-Crimson HcRed tdtomato AcGFP1 & DsRed-Monomer AcGFP1 & mcherry morange & mstrawberry morange & mcherry PAmCherry fluoresces when activated by 350 400 nm light. Dendra2 converts from green to red when exposed to UV or blue light. The bacterial expression of fluorescent proteins is driven by the lac promoter. Available vector formats allow targeting of fluorescent proteins to the golgi apparatus, actin filaments, plasma membrane, nuclei, microtubules, endoplasmic reticuli, mitochondria, peroxisomes, and endosomes (Figure 1). Red and far red fluorescent proteins are ideal for in vivo imaging due to their reduced autofluorescence. Our td Tomato fluorescent protein is very bright and ideal for in vivo imaging (Figure 2). FRET can be used to monitor the proximity and interaction of two proteins. Success of FRET relies on the correct pairing of donor and acceptor fluorescent proteins. Photoactivatable and photoconvertible fluorescent proteins are powerful tools for studying protein half-life and trafficking, because they allow the selective marking of molecules present at the time of activation/ conversion. A B A tdtomato B GFP Figure 1. Subcellular labeling. AcGFP1 and DsRed2 protein fusions were transiently transfected and visualized by fluorescence microscopy. Panel A. pacgfp1-mito (mitochondria) and pdsred2-nuc (nucleus) in HEK 293 cells. Panel B. pacgfp1-golgi (Golgi apparatus) and pdsred2-nuc (nucleus) in HEK 293 cells. Figure 2. In vivo Imaging. False-color overlay images (regions of interest encircled) highlight tdtomato fluorescence, but not GFP fluorescence, in the cadaver model. Panel A. Implanted tube with 100 x 10 6 MDA-MB-231-tdTomato-expressing cells, imaged with the DsRed filter set. Panel B. Implanted tube with 100 x 10 6 MDA-MB-231-GFP-expressing cells, imaged with the GFP filter set. Exposure times: 1 sec.

Antibodies Clontech offers highly specific antibodies for the detection of our Living Colors fluorescent proteins. Our antibodies are compatible with many applications, including Western blotting, immunoprecipitation, and immunocytochemistry. EGFP Monoclonal Full-Length ZsGreen Polyclonal A.v. Monoclonal (JL-8) Full-Length GFP Polyclonal A.v. Peptide GFP Monoclonal Proven Uses a WB, IP, IC WB WB, IP, IC WB WB, IP, IC WB, IP, IC AcGFP1 ZsGreen1 GFP GFPuv EGFP b EYFP b ECFP b a WB = Western Blot, IP = immunoprecipitation, IC = immunocytochemistry b Not sold by Clontech Selection Guide for Living Colors Fluorescent Proteins DsRed Monoclonal DsRed Polyclonal mcherry Monoclonal Anti-RCFP Polyclonal Pan Proven Uses a WB WB, IP, IC WB WB mplum E2-Crimson mraspberry HcRed1 mcherry mstrawberry PAmCherry AsRed2 DsRed-Monomer DsRed2 tdtomato DsRed-Express DsRed-Express2 morange morange2 mbanana ZsYellow1 Timer ZsGreen1 AmCyan1

CherryPicker Cell Labeling & Capture CherryPicker systems provide a powerful sample preparation tool for monitoring and capturing a homogeneous population of cells that either: A) exhibit a specific promoter activity, or B) express a specific protein of interest Benefits of CherryPicker A universal, low-background magnetic bead-based cell capture system. Ideal for expression profiling, marker discovery, and posttranslational modification applications. Gentle enrichment of a homogeneous cell population under sterile conditions (unlike FACS). Simple and convenient bench-top protocol. How does CherryPicker work? The system utilizes a chimeric membrane-anchored fluorescent protein, CherryPicker, which can be monitored via fluorescence microscopy, and captured on magnetic beads via a specific antibody. CherryPicker is only expressed under the control of your promoter of interest, or together with your protein of interest via a bicistronic IRES expression cassette (Figure 3). Therefore, only the cells with your activity of interest will be enriched (Figure 4). A P CMV Enrich cells expressing your protein of interest Your gene IRES CherryPicker gene CherryPicker protein CherryPicker antibody Your protein kd 55 34 Untransfected 35 x 103 cells 52.5 x 103 cells Before capture 35 x 103 cells 17.5 x 103 cells After capture Phospho-p44/42 MAPK (Erk 1/2) Magnetic beads 1 2 3 4 5 B Enrich cells with your active promoter GAPDH TF CherryPicker gene Your promoter Expression Binding Capture Figure 3. CherryPicker System. Panel A. Insert your gene of interest into the IRES reporter vector. Your protein will be expressed with CherryPicker. Panel B. Insert your promoter of interest into the promoterless reporter vector. CherryPicker will be expressed under the control of your promoter. CherryPicker is targeted to the membrane, where it can be visualized by microscopy, and/or captured via magnetic beads. Figure 4. Enrichment of MEK-CA-expressing cells via CherryPicker Systems. HEK-293 cells were transiently transfected with the IRES-CherryPicker expression vector encoding a constitutively active mutant of MEK (MEK-CA). Transfected cells were mixed 1:10 with untransfected cells. The mixed (Lanes 2, 3 & 4) and untransfected (Lane 1) cell lysates were analyzed by Western blot before and after capture. Western blot analysis was performed using a phosphospecific antibody against phosphorylated ERK1/2. GAPDH served as a loading control for total protein. Cat. # Product Package Size 632570 632571 CherryPicker Assay Kit 60 rxns 120 rxns 632572 CherryPicker Cell Capture System 60 rxns 632573 CherryPicker Cell Capture System (IRES) 60 rxns 632574 Lenti-X CherryPicker Cell Capture System 60 rxns 632575 Lenti-X CherryPicker Cell Capture System (IRES) 60 rxns

Visit www.clontech.com for a complete vector list. Cat. # Product Package Size Bacterial Expression 632468 pacgfp1 Vector 20 μg 632440 pamcyan Vector 20 μg 632412 pdsred-express Vector 20 μg 632535 pdsred-express2 Vector 20 μg 632553 pe2-crimson Vector 20 μg Subcellular Labeling 632591 ppamcherry-mito Vector 10 μg 632488 pacgfp1-tubulin Vector 20 μg 632583 pautophagsense Vector 20 μg 632409 pdsred2-er Vector 20 μg 632464 pacgfp1-golgi Vector 20 μg In Vivo Imaging 632531 ptd Tomato Vector 20 μg 632527 pmplum Vector 20 μg 632410 phcred1 Vector 20 μg 632553 pe2-crimson Vector 20 μg 632446 pzsgreen Vector 20 μg FRET 632470 pacgfp1-c1 Vector 20 μg 632466 pdsred-monomer-c1 Vector 20 μg Protein Dynamics & Half-Life 632545 pdendra2-n Vector 20 μg 632546 pdendra2-c Vector 20 μg 632584 ppamcherry-n1 Vector 10 μg 632585 ppamcherry-c1 Vector 10 μg Antibodies 632393 632392 Living Colors DsRed Monoclonal 20 μl 200 μl 632496 Living Colors DsRed Polyclonal 100 μl 632543 Living Colors mcherry Monoclonal 100 μl 632475 Living Colors Anti-RCFP Polyclonal Pan 100 μl 632377 632376 Living Colors A.v. Peptide 200 μl 1 ml 632569 Living Colors EGFP Monoclonal 100 μl 632375 Living Colors GFP Monoclonal 100 μl 632474 Living Colors Full-Length ZsGreen Polyclonal 100 μl 632380 632381 Living Colors A.v. Monoclonal (JL-8) 20 μl 200 μl 632593 632592 Living Colors Full-Length GFP Polyclonal 20 μl 100 μl 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 http://www.clontech.com. It is your responsibility to review, understand and adhere to any restrictions imposed by such statements. Takara Bio Europe SAS Clontech Laboratories www.clontech.com orders@clontech-europe.com tech@clontech-europe.com Europe : +33.(0)1 3904 6880 Austria : 0800 296 141 Germany : 0800 182 5178 Switzerland : 0800 563 629 United Kingdom : 0808 234 8063 For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resale. Clontech, the Clontech logo, AutophagSENSE, CherryPicker, Lenti-X, and Living Colors 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. 2012 Clontech Laboratories, Inc. This brochure is printed on 60% recycled paper. ZZXBR633398EU