Company introduction CYANAGEN Reagents Bio-Technology w.cyanage co n. m

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2 Company introduction CYANAGEN is a company based in Bologna (Italy), dedicated to the Research, Synthesis, Development, Production of Reagents for the research in the Bio-Technology field

3 Company History CYANAGEN was founded in 2003, Research Projects were its main core business until 2009 when a major company rearrangement occurred. Production and Marketing were then started

4 Company Association CYANAGEN is now part of a Group Of six Companies One of which HI TECH CYANAGEN is a controlled company

5 ISO Certification CYANAGEN is Quality Certified since 2010 ISO 9001:2008

6 Credit Certification CYANAGEN is Credit Certified since 2011

7 Credit Certification

8 Company Staff CYANAGEN 13 HI TECH CYANAGEN 2 MANAGEMENT 2 UNIBO CONTRACT 1

9 Global Market CYANAGEN is implementing a Worldwide Network of Dealers EUROPE BRAZIL ISRAEL SOUTH AFRICA INDIA JAPAN SOUTH KOREA TAIWAN CHINA

10 Sales Strategy CYANAGEN BRAND OEM APPLICHEM GEHC NACALAI PERKINELMER SERVA TAKARA

11 Product Lines Chemiluminescent substrates Protein assay kits Bioconjugation WESTAR for Western blot ELISTAR For ELISA Crosslinking Biotinylation Fluorescent Probes & Labels Fluorescent dyes Metal complexes CHROMIS DYES CHROMIS D+A CHROMIS DGE RuBy Protein Gel Stain Ruthenium Dyes

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13 WESTAR for Western blotting Luminol/enhancers solution Peroxide solution Protein detection complex + + =

14 WESTAR FEATURES Proprietary Technology Patented also in China Outstanding Sensitivity Wide Linear Dynamic Range Signal Stability Wide Range Product Line

15 WESTAR Selection Guide WESTAR ECL-SUN NOVA 2011 ETA C ETA C ULTRA SUPERNOVA Protein amount High High Medium Medium Very-low Detection limit ~ 20 pg ~ 2 pg ~ 500 fg ~ 150 fg ~ 50 fg 1 Ab dilution* 2 Ab dilution* 1:100 /1:5,000 1:1,000 / 1:15,000 1:500 / 1:5,000 1:20,000 / 1:100,000 1:1,000 / 1:15,000 1:25,000 / 1:150,000 1:5,000 / 1:50,000 1:50,000 / 1:250,000 1/5000 to 1/100,000 1/100,000 to 1/500,000 * Starting from 1 mg/ml of antibody stock solution

16 Competitors WESTAR Selection Guide Entry Level WESTAR ECL-SUN NOVA 2011 Amersham ECL- GEHC SUPER SIGNAL WEST PICO - PIERCE Amersham ECL START GEHC Amersham ECL- GEHC LUMI GLO RESERVE - KPL ECL PIERCE LUMINATA CLASSICO MILLIPORE LITE ABLOT PLUS EUROCLONE LUMI LIGHT - KPL WESTERN LIGHTNING PLUS - PERKINELMER LUMINATA CRESCENDO MILLIPORE BEYOECL PLUS - BEYOTIME WESTERN SURE ULTRA LICOR EZ-ECL-BIOLOGICAL INDUSTRIES

17 Competitors WESTAR Selection guide Mid Level WESTAR ETA C ETA C UTRA ECL PRIME-GEHC ECL PRIME - GEHC SUPER SIGNAL WEST DURA - PIERCE CLARITY - BIORAD LITE ABLOT EXTEND EUROCLONE ECL PLUS PIERCE LUMI LIGHT PLUS - KPL LITE ABLOT EXTEND EUROCLONE WESTERN LIGHTNING PRO - PERKINELMER LUMI LIGHT PLUS - KPL LUMINATA FORTE - MILLIPORE WESTERNBRIGHT ECL - ADVANSTA WESTERNSURE PREMIUM LICOR LUMINATA CRESCENDO MILLIPORE VISIGLO PRIME - AMRESCO LUMIGEN PS-3 LUMIGEN WESTERN BRIGHT QUANTUM - ADVANSTA

18 Competitors WESTAR Selection Guide Top Level WESTAR SUPERNOVA ECL SELECT GEHC SUPERSIGNALWESTFEMTO - PIERCE LUMI GLO ULTRA - KPL WESTERN LIGHTNING ULTRA - PERKINELMER LUMINATA FORTE MILLIPORE WESTERNBRIGHTSIRIUS - ADVANSTA FEMTOLUCENT G - BIOSCIENCES

19 WESTAR ECL-SUN 1 Ab: 1/100 1/5,000 2 Ab: 1/1,000 1/15,000 Detection level: ~ 20 picograms Direct alternative to the Pierce/Amersham ECL

20 WESTAR ECL-SUN Features High signal/noise ratio Working solution stable for at least 5 days (when properly stored in amber NALGENE bottle at 4 C) Stable signal emission

21 WESTAR ECL-SUN 1 Ab 1/1,000-2 Ab 1/15,000 [Transferrin] 56 ng 28 ng 14 ng 7 ng 3 ng 2 ng WESTAR ECL-SUN (Cyanagen) ECL Amersham (GEHC) ECL Pierce (Pierce Thermo Scientific) Luminata Classico (Millipore) LumiGLO (KPL)

22 WESTAR ECL-SUN Membrane Background ECL-SUN vs ECL Pierce

23 WESTAR ECL-SUN Background Comparison ECL-SUN (Cyanagen) ECL Pierce (Pierce Thermo Scientific) Exposure time: 51 minutes

24 WESTAR ECL-SUN Signal Stability ECL-SUN vs ECL Amersham

25 Signal Stability over time ECL-SUN (Cyanagen) Initial signal ECL Amersham (GEHC)

26 Signal Stability over time After 30 minutes ECL Amersham (GEHC)

27 Signal Stability over time After 1 hour ECL Amersham (GEHC)

28 Signal Stability over time After 2 hours ECL Amersham (GEHC)

29 WESTAR ECL-SUN Working Solution Stability ECL-SUN vs ECL Amersham/Pierce

30 Working Solution Stability over time ECL Amersham and Pierce WESTAR ECL-SUN Only 1 hour Low signals (analysis) reproducibility At least 5 days High signals (analysis) reproducibility

31 Working Solution Stability over time This is granted if the product is: Stored properly in amber NALGENE bottle at 4 C Kept away from direct light Not sized Not contaminated Not affected by additives WESTAR ECL-SUN At least 5 days High signals (analysis) reproducibility

32 WESTAR NOVA Ab: 1/500 1/5,000 2 Ab: 1/20,000 1/100,000 Detection level: ~ 2 picograms Alternative to the Pierce/Amersham ECL

33 WESTAR NOVA Ab 1/2,000-2 Ab 1/30,000 [Transferrin] 56 ng 28 ng 14 ng 7 ng 3 ng 2 ng WESTAR NOVA 2011 (Cyanagen) SuperSignal Pico (Pierce Thermo Scientific) Pico Tect (Imgenex)

34 WESTAR ETA C 1 Ab: 1/1,000 1/15,000 2 Ab: 1/25,000 1/150,000 Detection level: ~ 500 femtograms For medium-abundance proteins

35 WESTAR ETA C 1 Ab 1/4,000-2 Ab 1/40,000 [Transferrin] 56 ng 28 ng 14 ng 7 ng 3 ng 2 ng WESTAR ETA C (Cyanagen) ECL Plus/PS-3 (Pierce Thermo Scientific) Luminata Crescendo (Millipore) Clarity (BIORAD)

36 WESTAR ETA C ULTRA 1 Ab: 1/5,000 1/50,000 2 Ab: 1/50,000 1/250,000 Detection level: ~ 150 femtograms Extended signal duration for long exposure

37 WESTAR ETA C ULTRA 1 Ab 1/20,000-2 Ab 1/100,000 [Transferrin] 56 ng 28 ng 14 ng 7 ng 3 ng 2 ng WESTAR ETA C ULTRA (Cyanagen) Luminata Forte (Millipore) ECL Prime (GEHC) Visiglo Prime (Amresco) SuperSignal Dura (Pierce Thermo Scientific)

38 WESTAR SUPERNOVA 1 Ab: 1/5,000 1/100,000 2 Ab: 1/100,000 1/500,000 Detection level: ~ 50 femtograms The most sensitive substrate for high-end blotting

39 WESTAR SUPERNOVA 1 Ab 1/30,000-2 Ab 1/200,000 [Transferrin] 56 ng 28 ng 14 ng 7 ng 3 ng 2 ng WESTAR Supernova (Cyanagen) SuperSignal Femto (Pierce Thermo Scientific) ECL Select (GEHC) FemtoMAX (Rockland) LumiGLO Ultra (KPL)

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41 ELISTAR for ELISA CHEMILUMINESCENT SUBSTRATES ELISTAR Signal intensity Protein quantity Detection limit ETAC ULTRA ELISA SUPERNOVA ELISA Very High Extreme Low Abundance Very-low Abundance ~ 150 femtograms ~ 50 femtograms CYANAGEN PATENTED TECHNOLOGY ALSO IN CHINA

42 ELISTAR FEATURES More Sensitive Detection than Colorimetric ELISA; Higher Sensitivity and antibodies Saving; Immediate Light Generation with intense signal at both RT and 37 C; Wide Linear Dynamic Range - higher analysis reproducibility than Colorimetric (less tests required). Multiplex ELISA with ELISTAR

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44 QPRO Protein Quantitation The apple-green working reagent turns to purple reacting with proteins

45 QPRO QPRO and μqpro kits - based on BCA for rapid and sensitive detection of total protein concentration Product QPRO Standard mqpro Working Range μg/ml 0,5 20 μg/ml

46 QPRO FEATURES High Sensitivity Less Subject to Interferences from Detergents as Triton-X and SDS (1%) Linear Working Range 2 Year Shelf Life at RT Suitable to Microplates format

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48 CHROMIS DYES Different basic structures with precise Absorption and Emission to cover the Full Working Range Of Wavelength ( nm)

49 CHROMIS STRUCTURES CYANINES BODIPY 49/10

50 CHROMIS DYES FEATURES Each Fluorescent Compound is available with Different Charge (Anionic, Cationic, Zwitterionic and Neutral); Many different Reactive Moieties. Easy Selection of the optimal CHROMIS Dye for your application

51 CHROMIS DYES Selection Table Ex-Max Em-Max Molar absorbance Quantum yield CHROMIS nm 476 nm 37,000 M -1 cm -1 n.a. CHROMIS nm 506 nm 88,000 M -1 cm CHROMIS nm 539 nm 50,000 M -1 cm CHROMIS nm 561 nm 54,000 M -1 cm CHROMIS nm 567 nm 150,000 M -1 cm CHROMIS nm 564 nm 77,000 M -1 cm CHROMIS nm 612 nm 50,000 M -1 cm CHROMIS nm 592 nm 60,000 M -1 cm CHROMIS nm 605 nm 75,000 M -1 cm CHROMIS nm 624 nm 50,000 M -1 cm CHROMIS nm 646 nm 113,000 M -1 cm CHROMIS nm 663 nm 250,000 M -1 cm CHROMIS nm 707 nm 240,000 M -1 cm CHROMIS nm 790 nm 270,000 M -1 cm CHROMIS nm 840 nm 250,000 M -1 cm CHROMIS nm 851 nm 200,000 M -1 cm

52 Commercial Cyanine Structures FEATURES: High Extintion Molar Coefficient Mark any M.W. compounds : proteins, antibodies, nucleic acids. Applications: Flow Citometry, Western Blotting, Microscopy, Imaging

53 CHROMIS GEHC (Cy3, Cy5) Most used Cyanines from GEHC GEHC CYANINE l Abs (nm) l Em (nm) QY e (Mol -1 cm -1 ) CYANAGEN CYANINE Cy , CHROMIS 550 A Cy , CHROMIS 590 A Cy , CHROMIS 645 A Cy , CHROMIS 678 A Each Cyanagen s Cyanine is available with: Different Charge values (Anionic, Cationic, Zwitterionic and Neutral); Many different Reactive Moieties. WORK IN PROGRESS: new Cyanines with higher brightness, comparable with Alexa Fluor dyes!

54 CHROMIS: NIR Cyanine Dye - Cy7 CYANAGEN CYANINE Abs (nm) Em (nm) e (Mol -1 cm -1 ) QY Competitors CHROMIS ,1 IR Dye 800 CW CHROMIS ,1 CHROMIS ,1 FEATURES: Strong tissue penetration: ideal for in vivo Imaging Minimal autofluorescence in the IR region: Undetectable noise High Brightness and Photostability for WB, Imaging and FC WORK IN PROGRESS: new Cyanines with higher brightness

55 Cyanine: LSS Dyes Large Stokes Shift: Cyanines with Abs of Cy5/ Cy5.5 and Emission in IR region CYANAGEN LSS Dyes Abs (nm) Em (nm) e (Mol -1 cm -1 ) Laser CHROMIS He-Ne CHROMIS He-Ne

56 Commercial Bodipy Structure CHROMIS 500 N FEATURES: High Quantum Yield Mark low M.W. compounds: aminoacids, nucleotides, (NO Ab) Unique hydrophobic properties, ideal for staining lipids, membranes, and other lipophilic compounds

57 CHROMIS: BODIPY Dyes CYANAGEN BODIPY l Abs (nm) l Em (nm) QY e (Mol -1 cm -1 ) COMPETITORS CHROMIS 500 N BODIPY FL CHROMIS 515 N , BODIPY FL CHROMIS 530 N , BODIPY R6G CHROMIS 555 N , BODIPY 558/568 CHROMIS 570 N , BODIPY 576/589 CHROMIS 630 N , BODIPY 630/650 WORK IN PROGRESS: new CHROMIS 500 water soluble as alternative to FITC and ALEXA FLUOR 488!

58 Ru COMPLEXES d 6 COMPLEXES of Ru 58/10

59 Ru COMPLEXES l abs : 432 nm l em : 618 nm FEATURES: Unique Features different from common organic chromophores; Singular spectrum of absorption/emission with a large Stokes Shift; Long Duration of light emission; High Chemical and Photochemical Stability of labeled compounds.

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61 CHROMIS D+A Protected by US and their foreign equivalents and pending patents

62 CHROMIS D+A FEATURES Easy: the reaction is carried out by mixing the supplied reagents and adding the target Ab; Fast: no weighing of tiny amounts of activated dye is needed; Efficient: the reactive dye is quantitatively generated in situ, thus affording the maximum efficiency.

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64 Protein Separation Methods SDS-PAGE Few dozens of proteins bands Separate proteins according to their electrophoretic mobility and no other physical feature 2-D Electrophoresis Thousand of proteins bands Complex mixtures of proteins are separated in two dimensions by isoelectric point and protein mass Difference Gel Electrophoresis Thousand of proteins bands with more accuracy Detection of the smallest possible differences in protein expression with superior statistical confidence

65 2D-DGE FEATURES Size and charge matched: labeled samples co-migrate with gel; Bright highly sensitive: enable minimal labeling technique; Photo stable: minimal loss of signal; ph insensitive: no change in signal over wide ph.

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67 Crosslinker/PEGylation/Biotinylation Wide range of products that can be supplied on demand

68 Large Stock of the following products is available Homofunctional BS3 DSG DSS DTSP EGS Orders from 1g. CROSSLINKERS for Bio-conjugation For other products of the same line special agreements can be provided from 1 g LEADTIME To Be Defined

69 Large Stock of the following products is available Heterofunctional GMBS MBS SIA SATA Orders from 1 g. CROSSLINKERS for Bio-conjugation For other products of the same line special agreements can be provided from 1 g LEADTIME To Be Defined

70 SERVICES Design and Synthesis of Fluorescent Dyes according to Client s specifications; Customized R&D programs in the diagnostic and bioanalytical fields; National and International Collaborations for research calls; Technical support; Training, seminars, webinars, wet workshops.

71 Webinars and Client Support

72 Coming Soon Products

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74 Ruby Gel Stain - Protein Quantitation Features: Ready to Use kit; Improved Signal/Background Ratio; Fast Staining in less than three hours; Same Sensitivity of Silver-staining without its drawbacks; Good Linearity and Homogeneity.

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76 Green Stain DNA Quantitation FEATURES: NON TOXIC SENSITIVE Detects 250 pg of DNA per band HIGH CONTRAST Bright Green Fluorescence with exceptionally low background

77 Green Stain DNA Quantitation CYANAGEN GREEN STAIN COMPETITORS SyBr Safe, Gel Red, Gel Green, EtBr, SyBr Green 1 kb Plus DNA Ladder (Invitrogen, no ) on a 1% agarose Lane 1 5 ng/lane Lane 2 10 ng/lane Lane 3 20 ng/lane

78 Green Stain DNA Quantitation EASY TO USE Substitutes Ethidium Bromide in Molten Agarose. Can be used in different staining protocols: gel staining after electrophoresis, gel pre-staining, sample pre-staining. The detection can be performed with: standard UV transilluminator; CCD-based image documentation systems.

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80 RENEW - Stripping Buffer Pierce Restore After stripping Cyanagen Renew Pierce Restore Cyanagen Renew Reprobing FEATURES: Costs/Time saving: no re-run of gels and blots; re-probe membrane with the same sample; Effective: more efficient than homemade buffers; Gentle: not damaging Ab; Odorless: no mercaptans.

81 Thank you for your kind attention