+1 530 888 530 8925 +1 Fax 130-097-256 130-097-257 130-097-259 CA 95602, USA magnetically labeled +1 530 888 8871, 888 530 8871, +1 , Elute -cell complexes in cell cells. culture medium and culture Magnetically label virus with with virus label Magnetically MACSductin Reagent. Mix MACSductin labeled virus with magnetically labeled cells. Apply labeled virus and cells to column. 800 FOR MACS Lindbergh Street, Auburn, Auburn, Lindbergh Street, Miltenyi Biotec Inc. Phone [email protected] 2303 genetic modification of primary human and murine cell and primaryhuman of modification genetic MACSductin Reagent virus preparations – no viral concentration required. viral – no concentration preparations subpopulations. Product applications Product Ex vivo virus titer low cells with of target transduction Effective cells. target of infection Synchronized Legend

Principle of magnetic transduction with MACS Technology. MACS with transduction magnetic of 1: Principle Figure primary murine and human target cells were demonstrated compared compared demonstrated were cells target human and primary murine alone.³ Magnetofection methods or transduction standard to the MACS Column leading to reduced viral reagent requirements requirements viral reagent reduced to leading Column the MACS an efficient colocalisation within the high-gradient magnetic field of field magnetic within high-gradient the colocalisation efficient an cells. magnetically only the labeled of target specific and transduction magselectofection, superior viral transduction efficiencies of both of efficiencies viral transduction superior magselectofection, Magnetic labeling of both virus particles and the target cells ensures ensures cells both the virus target of particles and labeling Magnetic ● ● ● 0.5 mL 3×0.5 mL 0.25 mL MACSductin™ Reagent MACSductin™ 1.2 page 1/4

or Bergisch Gladbach,Bergisch 10⁹ cells +49 2204 85197 × ×10⁸ cells Fax 68, 51429 68, 51429 130-097-257) or 10⁷–3 8 °C. Do not freeze. Do°C. 8 not × 10⁷–10⁹ cells or 10⁷–10⁹ cells − × +49 2204 8306-0, transduction of6 Miltenyi Biotec GmbH Friedrich-Ebert-Straße Phone [email protected] [email protected] Germany MACSductin Reagent (# 130-097-259) Reagent MACSductin

×0.5 mL for ×0.5 mL solution without preservatives. MACSductin™ Reagent (# 130-097-256) or Reagent 0.25 mL MACSductin™ (# Reagent 0.5 mL MACSductin 3 3 0.5 mL for transduction of 2 MACSductin Reagent is supplied in a sterile in a sterile supplied is Reagent MACSductin Store protected from light at 2 at light from protected Store for transduction of 10⁷–5 of transduction 0.25 mL for The expiration date is indicated on vialthe label. is indicated date expiration The

Combined magnetic cell viral and sorting transduction applications Product requirements instrument and Reagent requirements Calculation reagent of selected positively of cells target Transduction cell monocultures of Transduction Transduction of human cell lines with human adenoviral of vector Transduction

Reagent 2.1 1.1 1.2 1.3 2.2 2.3 Examples of magnetic transduction using MACSductin™ magnetic MACSductin™ Examples of transduction using 3.1 Optimization of transduction parameters Troubleshooting Combined magnetic cell viral and sorting transduction www.miltenyibiotec.com

References

is a method that enables cell modifying agents (e.g. RNA, DNA, DNA, RNA, (e.g. cell modifying agents enables method a is that in cell culture, the major factor limiting infection is the diffusion of the diffusion is infection limiting factor the major cellin culture, into a single process to significantly improve viral transduction viral transduction improve significantly to process a single into the virus particles to the cell surface.¹ As the half life of of in cell the half life the virus the As cell surface.¹ particles to high-gradient magnetic field on the column matrix. Magnetofection Magnetofection matrix. on column the field magnetic high-gradient Storage Size approximately 50 then Labeled are cells approximately in diameter. nanometers and Magnetofection®. MACS Technology is the gold standard in standard the gold is Technology MACS Magnetofection®. and surface using magnetic polycationic carriers.² polycationic magnetic surface using specifically retained within a MACS Column when it is placedis in a it when Column a MACS specificallyretained within maximize infection or transduction events. transduction or infection maximize format Product 5. culture is short (e.g. 5–8 hours for retroviruses)¹, it is important to to important is it retroviruses)¹, for 5–8 hours (e.g. short is culture to cells delivery ofthetarget virus tothe rapid and efficient an ensure cell separation. Target cells are specificallyantibody- labeled with are cells Target cell separation. MicroBeads, MACS nanoparticles, superparamagnetic conjugated efficiencies and improve sample handling: MACS® Technology MACS® handling: sample improve and efficiencies MACS Separator. This is a strong permanent magnet which induces a induces magnet which permanent strong is a This Separator. MACS MACSductin Reagent combines two established technologies technologies established two combines Reagent MACSductin 6. viruses) to be concentrated by magnetic force at the target cell’s cell’s the target at force magnetic by viruses) be to concentrated During the infection of target cells with native or recombinant viruses recombinant or native with cells target of During the infection 3. Components Protocol 2. Protocol Contents

4. When the two methods are combined into a single process, termed process, a single into combined methods the are two When 1. Description 1.1 1. Description

140-003-567.01 target ⁶ MACS Column MS MS LS LS MS MS LS LS 10 75 37.5 375 10 10 10 37.5 Minimum complex (µL) volume 2.5 25 12.5 125 0.25 2.5 1.25 12.5 MACSductin (µL) Reagent 1×10⁵ 1×10⁶ 5×10⁵ 5×10⁶ 1×10⁵ 1×10⁶ 5×10⁵ 5×10⁶ No. of infectious (IU) virions MOI 0.1 1 0.1 0.5 0.1 1 0.1 0.5 briefly centrifuge sample at 13,000×g for 30 seconds to remove remove to 30 seconds for at 13,000×g sample centrifuge briefly in the positive fraction. in the positive particulate matter. Transfer supernatant to a fresh tube, avoiding avoiding tube, fresh a to supernatant Transfer matter. particulate sterile filter (0.22 or 0.45 µm). (0.22 filter sterile retro- and lentiviral samples should be thawed rapidly at at rapidly be thawed should samples lentiviral and retro- cells at desired MOI. desired at cells floating fragments. Alternatively, filter through an appropriate appropriate an through filter Alternatively, fragments. floating (see “Before starting” above), determine the target cell number cell number the target determine above), starting” (see “Before Calculate amount of virus sample required to transduce target target transduce to required virus sample of Calculate amount Following positive selection of magnetically labeled target cells cells magnetically labeled selection target of positive Following Transduction of positively selected positively of cells target Transduction ice, on be should thawed Adenoviruses virus sample. Thaw 37 °C. If the virus supernatant has not been previously purified, purified, been previously not has 37 °C. thevirus supernatant If The virus aliquot should have a volume at least twice the volume volume least at twice the volume a have should virusThe aliquot

To accurately determine the absolute physical particle titer of the of particle titer physical the absolute determine accurately To Target cell number Unpurified supernatant 1×10⁶ 1×10⁶ 5×10⁶ 1×10⁷ Purified vector preparation Purified 1×10⁶ 1×10⁶ 5×10⁶ 1×10⁷ page 2/4 transduction using unpurified supernatant and purified vector preparation. purified vector and supernatant unpurified using transduction lentiviral vectors) or a DNA slot-blot approach⁴ (for adenoviral vectors). adenoviral (for approach⁴ slot-blot DNA a or vectors) lentiviral the infectious titer of the virus sample will only allow an approximate approximate an allow will only the virus sample of titer the infectious titering kit (e.g. virus-associated p24 ELISA or (RT-) PCR based for (RT-) virus-associated p24 ELISA or (e.g. kit titering purified lentiviral particles, the MACSductin Reagent requirement requirement Reagent MACSductin particles, the lentiviral purified are infectious (e.g. 1 in 20) and the physical particle number is often often is particle number the physical 1 in 20) and (e.g. infectious are supernatant, 25 µL of MACSductin Reagent is required. When using using When required. is Reagent MACSductin 25 µL of supernatant, 1. in table shown free medium before use. use. before free medium used as a measurement of viral titer. viral titer. of used a measurement as general rule of thumb, 1 in 1000 retro-/lentivirus vector particles in vector 1 in 1000 retro-/lentivirus rule thumb, general of retro-/lentivirus while in 100 1 infectious are supernatant unpurified estimation of the total virus particle titer to be calculated. As a be As to calculated. virus the total particle titer of estimation Reagent will complex both infectious and defective virus particles, defective and both infectious will complex Reagent Examples of MACSductin Reagent requirements for lentiviral vector vector lentiviral for requirements Reagent MACSductin of 1: Examples Table of MACSductin Reagent. If required, dilute virus aliquot with serum-with virus aliquot dilute required, If Reagent. MACSductin of cells at a multiplicity of infection of 1 (MOI=1) with unpurified unpurified with 1 (MOI=1) of infection of a multiplicity at cells further 2.5 µL. Some 10 to examples of a factor by becan reduced are transduction a lentiviral for amounts reagent calculate to how of For example, for a standard lentiviral transduction of 1×10 of transduction lentiviral standard a for example, For Generally, for purified adenoviral vectors more than 1 in 100 100 in 1 particles than more vectors adenoviral purified for Generally, vector particles in purified vector preparations are infectious. infectious. are preparations vector particlesin purified vector Column or 5 mL for transduction using an LS Column. an using transduction 5 mL for or Column volume used should not be less than 10 µL and maximal complex complex maximal 10 µL and be than less not used should volume MS an using transduction exceed 2 mL for not should volumes virus samples, it is recommended to use an appropriate virus particle appropriate use an to recommended is it virus samples, 2. 2.2 3. 1. For optimum performance, the minimum complex-forming complex-forming the minimum performance, optimum ▲ For ▲ ▲ are not not are 2+ or Mg or 2+ serum albumin, human serum, serum human albumin, MiniMACS, OctoMACS, OctoMACS, MiniMACS, QuadroMACS, MidiMACS, Separator VarioMACS, SuperMACS II VarioMACS, SuperMACS II VarioMACS,

progenitor cells, 10⁵ cells can also can cells be10⁵ cells, progenitor + 7.2, 0.5% bovine serum 7.2, 0.5% bovine albumin Max. number Max. number transduced 5×10⁶ of labeled be to cells 10⁷ transduced 2.5×10⁶ of labeled be to cells Min. number 10⁶ Column adapters are required to insert certain columns into the into insert to certain columns required are adapters Column EDTA can be replaced by other supplements such as anticoagulant anticoagulant as such supplements other by be can replaced EDTA For very rare cells, such as CD34 as such cells, very rare For

376) 1:20 with autoMACS® Rinsing Solution Solution Rinsing autoMACS® 130‑091‑376) 1:20 with Note: Note: Note: Note:

MS LS Column buffered pH saline (PBS), buffered labeled cells that are to be transduced and to the number of total cells. total of the number to and be to transduced are labeledthat cells leukaemia celllines. MACS Separator data sheet. data Separator MACS recommended for use. required supplements required MACS Separator and MACS Columns according to the number of the number to according Columns MACS and Separator MACS viruses. or fetal bovine serum (FBS). Buffers or media containing Ca mediaor containing serum fetal bovine (FBS). Buffers or efficiently transduced in an MS Column when using an MOI≥10. an using when an MS Column in transduced efficiently citrate dextrose formula-A (ACD-A) or citrate phosphate dextrose (CPD). BSA BSA (CPD). dextrose phosphate citrate or (ACD-A) formula-A dextrose citrate human as such proteins other by becan replaced °C). (2−8 cold buffer (# 130‑091‑222). Keep (BSA), and 2 mM EDTA by diluting MACS BSA Stock Solution Solution Stock BSA MACS diluting by EDTA 2 mM (BSA), and (# Reagent and instrument requirements instrument and Reagent Calculation of reagent requirements Calculation reagent of MACS Columns and MACS Separators: Choose the appropriate Choose the appropriate Separators: MACS and Columns MACS PEB buffer (degassed): Prepare a solution containing phosphate- containing a solution Prepare (degassed): buffer PEB Effective transduction of difficult-to-transduce target cells, e.g., cells, target difficult-to-transduce of transduction Effective wild-type and recombinant of a wide range with Compatible ▲ ▲ 1.5 mL, 15 mL, and 50 mL sterile tubes 50 mL sterile 1.5 mL, 15 mL, and MACS MultiStand (# 130-042-303) MultiStand MACS (# 130-042-401) LS Columns (# 130-042-201) or MS Columns RPMI 1640 (# 130-091-440) and e.g. medium, Cell culture Magnetically labeled target cell population enriched by MACS MACS by enriched cell population labeled target Magnetically Recombinant virus containing expression cassette of interest. of cassette expression virus containing Recombinant Technology. VarioMACS™ or SuperMACS™ II Separators. For details refer to the respective the respective to refer details For Separators. II SuperMACS™ or VarioMACS™ Protocol Before transducing cell monocultures, it is recommended to assess to recommended is it cell monocultures, transducing Before

by flow cytometry. For an overview of available reagents for use in thefor reagents overviewavailable an of For cytometry. flow by the presence of a suitable cell surface molecule for magnetic labeling magnetic cell surface molecule for a suitable of the presence populations prior to transduction as the presence of the MACSductin the MACSductin of the presence as transduction to prior populations selection efficiency. selection efficiency. fraction from a positive selection procedure, or they be can or a cell selection procedure, a positive fraction from magnetically labeled been with uniformly have that monoculture Before starting dependent on the ratio of MACSductin Reagent to virus to particles. Reagent MACSductin of the ratio on dependent Reagent within the MACS Separation Columns can reduce cell reduce can Columns Separation the MACS within Reagent field of hematology and immunology refer to www.miltenyibiotec.com. refer immunology and hematology of field MicroBeads. MACS MicroBeads. These cells can either comprise the enriched enriched the comprise either TheseMicroBeads. cells can MACS ● ● ● ● ● ● ● ● ● ● 2.1

▲

Unless otherwise specifically indicated, Miltenyi Biotec Biotec otherwiseMiltenyi specifically indicated, Unless for not and use only research services and for products are use. therapeutic or diagnostic 2. 1.3 The efficient formation of magnetic virus transduction complexes is complexes magnetic of transduction virus formation efficient The As not all virions in a virus sample are infectious and MACSductin MACSductin and infectious are in a virus all sample virions not As ▲ It is recommended to pre-enrich the target cells from mixed cell mixed from cells the target pre-enrich to recommended is ▲ It The target cell population must magneticallymust be labeled with cell population ▲ The target

140-002-817.01 4. Resuspend MACSductin Reagent by pipetting or vortexing. sterile filter (0.22 or 0.45 µm). Pipette required volume of MACSductin Reagent (refer to 2. Determine the target cell concentration in the cell sample. section 2.1) into a fresh tube. ▲ Note: If only small volumes of MACSductin Reagent are required, a diluted working 3. Calculate amount of virus sample required to transduce target stock can be prepared by diluting MACSductin Reagent in serum-free medium. cells at desired MOI. 5. Add the virus aliquot to the MACSductin Reagent and incubate 4. Transfer required number of target cells to a suitable tube and at room temperature for 20 minutes. pellet at 100–200×g for 10 minutes. Remove supernatant and ▲ Note: For optimum performance, the minimum complex-forming volume used resuspend cell pellet in 100 µL PEB per 10⁷ total cells (minimum should not be less than 10 µL and maximal complex volumes should not exceed 2 mL for transduction using an MS Column or 5 mL for transduction using an LS recommended volume 100 µL). Column. ▲ Note: The maximal labeled cell numbers should not exceed 5×10⁶ for ▲ Note: The virus aliquot should have a volume at least twice the volume of transduction using an MS Column or 1×10⁷ for transduction using an LS Column. MACSductin Reagent. If required, dilute virus aliquot with serum-free medium before use. 5. Add MicroBeads specific for a cell surface molecule on the target cell at the recommended titer. For details refer to the respective 6. Transfer required number of target cells to a suitable tube and data sheet. Mix well and incubate at 2–8 °C for 15 minutes. pellet at 100–200×g for 10 min. Remove supernatant. Resuspend cells in 500 µL or 1000 µL serum-free medium for transductions 6. During the cell labeling step, resuspend MACSductin Reagent using MS or LS Columns, respectively. by pipetting or vortexing and pipette required volume of ▲ Note: The maximal labeled cell numbers should not exceed 5×10⁶ for MACSductin Reagent (refer to section 2.1) into a fresh tube. transduction using an MS Column or 1×10⁷ for transduction using an LS Column. ▲ Note: If only small volumes of MACSductin Reagent are required, a diluted ▲ Note: For optimal performance it is recommended to use a minimum of 1×10⁶ working stock can be prepared by diluting MACSductin Reagent in serum-free cells for transduction using an MS Column or 5×10⁶ for transduction using an LS medium. Column. However, for very rare cells such as CD34+ progenitor cells, 1×10⁵ cells can also be efficiently transduced using an MS Column when at an MOI ≥10. 7. Add the virus aliquot to the MACSductin Reagent and incubate at room temperature for 20 minutes. 7. Place column in the magnetic field of a suitable MACS Separator. ▲ Note: The virus aliquot should have a volume at least twice the volume of For details refer to the respective MACS Column data sheet. MACSductin Reagent. If required, dilute virus aliquot with serum-free medium before use. 8. Prepare column by rinsing with appropriate amount of serum- ▲ Note: For optimum performance, the minimum complex-forming volume used free medium: should not be less than 10 µL and the maximal complex volumes should not exceed MS: 500 μL LS: 3 mL 2 mL for transduction using an MS Column or 5 mL for transduction using an LS Column. 9. Gently add the target cells to the virus-MACSductin Reagent complex and mix by pipetting up and down. Do not vortex. 8. After completion of magnetic cell labeling, add 10 volumes of serum-free medium to the target cells and pellet at 100–200×g 10. Place an empty tube under the column to collect waste and apply for 10 minutes. Remove supernatant. Resuspend cells in 500 µL the mixture of labeled virus and cells onto the column. Collect or 1000 µL serum-free medium for transduction using MS or LS flow-through fraction. Columns, respectively. 11. Wash column once with appropriate amount of complete culture 9. Place column in the magnetic field of a suitable MACS Separator. medium and collect flow-through in waste tube. For details refer to the respective MACS Column data sheet. MS: 1 mL LS: 3 mL 10. Prepare column by rinsing with appropriate amount of serum- 12. Remove column from the separator and place it on a suitable free medium: collection tube. MS: 500 μL LS: 3 mL 13. Pipette appropriate amount of complete medium onto the 11. Gently add the target cells to the virus-MACSductin Reagent column. Immediately flush out the virus-cell complexes by complex and mix by pipetting up and down. Do not vortex. firmly applying the plunger supplied with the column. MS: 1 mL LS: 3 mL 12. Place an empty tube under the column to collect waste and apply the mixture of labeled virus and cells onto the column. Collect 14. Transfer eluted cells without bubbles into a cell culture-plate at a flow-through fraction. suitable cell density. 13. Wash column once with appropriate amount of complete culture 15. Place cells in a cell culture incubator and cultivate under medium and collect flow-through in waste tube. standard conditions. MS: 1 mL LS: 3 mL

2.3 Transduction of cell monocultures 14. Remove column from the separator and place it on a suitable collection tube. Magnetically selected cells that have been cultured for a period of time or immortalised cell lines can also be transduced using the 15. Pipette appropriate amount of complete medium onto the MACSductin Reagent. In this case all cells are magnetically labeled column. Immediately flush out the virus-cell complexes by and transduced on the column. firmly applying the plunger supplied with the column. MS: 1 mL LS: 3 mL 1. Thaw virus sample. Adenoviruses should be thawed on ice, retro- and lentiviral samples should be thawed rapidly at 16. Transfer eluted cells without bubbles into a cell culture-plate at a 37 °C. If the virus supernatant has not been previously purified, suitable cell density. 140-002-817.01 briefly centrifuge sample at 13,000×g for 30 seconds to remove 17. Place cells in a cell culture incubator and cultivate under particulate matter. Transfer supernatant to a fresh tube, avoiding standard conditions. floating fragments. Alternatively, filter through an appropriate

Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for diagnostic or therapeutic use. page 3/4 3. Examples of magnetic transduction using 5. Troubleshooting MACSductin™ Reagent Poor transduction efficiencies ▲ 3.1 Transduction of human cell lines with purified adenoviral Virus sample contains large number of defective, non-infectious vector particles. The absolute physical number of virus particles should be calculated by use of a commercial virus particle titering kit. For retroviral and lentiviral vectors these can either be ELISA-based kits A B (e.g. virus-associated p24 ELISA) or (RT-)PCR based (e.g. viral RNA 100 3000 18 50 RT-PCR quantitation kit). For adenoviral vectors a DNA slot-blot

90 16 45 approach can be used.⁴ 2500 80 14 40 ▲ Decay of virus-MACSductin Reagent complexes. Only freshly 70 35 2000 12 prepared complexes should be used. Storage of the complexes for 60 30 more than 1 hour can reduce transduction performance. 10 50 1500 25 ▲ 8 Too few target cells were used. A minimum of 1×10⁶ target cells 40 20 should be immobilized on MS Columns and 2.5×10⁶ cells on LS 1000 6 30 GFP MFI 15 GFP MFI Columns. Using lower cell densities may lead to a reduction in 20 4 10 + 500 transduction efficiency. However, for very rare cells such as CD34 % GFP-positive cells % GFP-positive cells % GFP-positive 10 2 5 progenitor cells, 1×10⁵ cells can also be efficiently transduced with 0 0 0 0 lentiviral vectors using an MS Column at MOI ≥10. A549 A549 and HuT78 HuT78 and MACSductin MACSductin ▲ Too many target cells were used. A maximum of 5×10⁶ target cells should be immobilized on MS Columns and 1×10⁷ cells on LS %GFP+ cells GFP MFI (mean fluorescence intensity) Columns. Exceeding these cell densities may lead to a reduction in transduction efficiency. For larger cell numbers, use a number of Figure 2: A large increase in gene marking efficiency is observed after adenoviral columns in parallel. transduction of human cell lines with MACSductin Reagent. 1×10⁶ HuT78 and A549 cells were magnetically labeled with CD30 or CD44 MicroBeads, Poor cell recoveries respectively. In parallel purified adenoviral vector particles encoding GFP were ▲ Insufficient magnetic labeling. The surface marker used for complexed with MACSductin Reagent. The A549 cells were then transduced in MS retention of the cells on the separation column must be suitable Columns at a pMOI (physical particles per cell) of 50 while HuT78 cells were transduced for magnetic separation and should be sufficiently expressed. Some at a pMOI of 500. GFP expression from transduced cells was increased up to 30-fold 24 hours post-transduction as measured by flow cytometry (data courtesy of Dr. Florian cell surface markers can be downregulated when cells are cultivated Kreppel and Dr. Sigrid Espenlaub, Department of , University of Ulm). at high density. Surface marker expression can be verified by flow cytometric analysis. For more information about MACS Products for cell separation refer to www.miltenyibiotec.com, for MACSmolecular products and Reduced cell viability ▲ services refer to www.macsmolecular.com. Amount of viral vector or MACSductin Reagent used is too high. Transduction of cells with excess viral vector can lead to cellular toxicity. Also some cells may react sensitively to large amounts 4. Optimization of transduction parameters MACSductin Reagent. In these cases it is recommended to reduce Magnetic transduction of target cells with MACSductin Reagent is the MOI applied or the amount of MACSductin Reagent used. a reliable and robust technique. However, the quality of the virus preparation as well as the phenotype and growth characteristics of the 6. References target cells can affect transduction efficiency (refer also to section 2.1). 1. Andreadis, S. et al. (2000) Toward a more accurate quantitation of the activity of To further optimize transduction parameters, it is recommended to recombinant retroviruses: alternatives to titer and multiplicity of infection. J. Virol. titrate the amount of MACSductin Reagent using a fixed cell number 74: 1258–1266. and fixed MOI. Examples using different viral vector preparations 2. Plank, C. et al. (2003) The magnetofection method: using magnetic force to with 1×10⁶ target cells are shown below. enhance . Biol. Chem. 384: 737–47. 3. Sanchez-Antequera, Y. et al. (2011) Magselectofection: an integrated method of ▲ For unpurified lentiviral vector supernatant it is recommended to nanomagnetic separation and genetic modification of target cells. Blood 117: e171–181. use 6, 12, 25, 50, and 100 μL MACSductin Reagent for 1×10⁶ cells, 4. Kreppel, F. et al. (2002) A DNA-based method to assay total and infectious particle MOI=1. contents and helper virus contamination in high-capacity adenoviral vector preparations. Human Gene Therapy 13: 1151–1156. ▲ For purified lentiviral vector preparations it is recommended to use 0.6, 1.2, 2.5, 5, and 10 μL MACSductin Reagent for 1×10⁶ cells, All protocols and data sheets are available at www.miltenyibiotec.com. MOI=1. Warranty ▲ For purified adenoviral vector preparations it is recommended to The products sold hereunder are warranted only to be free from defects in workmanship use 1.25, 2.5, 5, 10, and 20 μL MACSductin Reagent for 1×10⁶ cells, and material at the time of delivery to the customer. Miltenyi Biotec GmbH makes pMOI=500 (pMOI=physical particles per cell) no warranty or representation, either expressed or implied, with respect to the fitness of a product for a particular purpose. There are no warranties, expressed or implied, which extend beyond the technical specifications of the products. Miltenyi Biotec GmbH’s liability is limited to either replacement of the products or refund of the purchase price. Miltenyi Biotec GmbH is not liable for any property damage, personal injury or economic loss caused by the product. autoMACS, MACS, and MACSQuant are registered trademarks and MACSductin, MidiMACS, MiniMACS, OctoMACS, QuadroMACS, SuperMACS, and VarioMACS 140-002-817.01 are trademarks of Miltenyi Biotec GmbH. Magnetofection is a registered trademark of Dr. Christian Plank and Christian Bergemann. Copyright © 2012 Miltenyi Biotec GmbH. All rights reserved.

Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for diagnostic or therapeutic use. page 4/4