140-001-861.06 1. Warnings These precautions are recommended to avoid deposits in plumbing in plumbing to avoiddeposits are recommended precautions These 1. Description 4. This product is for research use only.use is forresearch product This The CD154 has been tested to cross-react with rhesus rhesus with cross-react to tested been has CD154The antibody

Contents 2. Capacity Components 3. fascicularis where explosive conditions may develop. may conditions explosive where Reagents contain sodium azide. Under acidic conditions sodium sodium conditions Under acidic azide. sodium contain Reagents Product format discarding. before water running with diluted be should compounds monkey ( monkey macsde www.miltenyibiotec.com www.miltenyibiotec.com azide yields hydrazoic acid, which is extremely toxic. Azide Azide toxic. extremely is which acid, hydrazoic yields azide Storage Phone Phone Friedrich-Ebert-Straße Friedrich-Ebert-Straße Miltenyi Biotec B.V. KG &Co. Biotec Miltenyi

Protocols Description Reference Example of a separation using the CD154 MicroBead Kit CD154 MicroBead the using of aseparation Example

2.3 2.2 2.1 1.4 Applications 1.3 1.2 1.1 +49 2204 8306-0, 2204 +49 @

miltenyi.com

CD154 expression Magnetic separation Magnetic labeling Magnetic 2.1.1 preparation Sample requirements instrument and Reagent information Background Separation MACS® of the Principle Macaca mulatta ). In vitro In vitro

at 2−8 from Store protected separations. up100 to cells, For 10⁹ total UltraPure: MicroBeads Anti-Biotin mL 2 All reagents are supplied in buffer containing containing buffer in supplied are reagents All human: CD154-Biotin, 1 mL UltraPure MicroBeads conjugated to to conjugated MicroBeads UltraPure Monoclonal anti-human CD154 antibody CD154 antibody anti-human Monoclonal IgG2a). conjugated to biotin (clone: 5C8, isotype: mouse mouse isotype: (clone: 5C8, biotin to conjugated mouse IgG1).mouse (isotype: antibody anti-biotin monoclonal indicated on the vial label. vial on the indicated is date expiration The freeze. azide. 0.05% sodium and stabilizer 68, 5142968, Fax Fax stimulation for induction of for induction stimulation +49 2204 85197 2204 +49 ) and cynomolgus monkey ( monkey cynomolgus ) and Bergisch Gladbach, Germany Bergisch Gladbach,

°C. Do not Do °C.

Macaca

1.4 Applications 1.3 1.2 1.1 T The antibody specifically recognizes the human CD154 human the recognizes specifically antibody The

human CD154 Kit MicroBead ● ● ● ● CD40L, gp39, T-BAM, TRAP, or Ly-62. CD154 is transiently gp39, T-BAM, CD154 or Ly-62. TRAP, transiently CD40L, is CD154 CD154 can be used as a marker for activated -specific CD4 antigen-specific for activated a marker as used be CD154 can in vitro in First, the CD154 the First, enriched T cells is used. is Tcells enriched of population apure if not required is of CD40 Blocking cells. of CD154 expression down-regulation prevents suspensions of cell containing the CD the containing role as a costimulatory molecule in /antigen-presenting cell cell Tcell/antigen-presenting in molecule acostimulatory role as CD154 retained magnetically the field, magnetic the from column the removing of CD154 depleted thus is fraction cell this through; run up-regulated on activated CD4 fraction. fraction. suspension is loaded onto a MACS® Column, which is placed in in placed is which Column, onto a MACS® loaded is suspension antigen, a 39 kDa transmembrane glycoprotein also known as as known also glycoprotein transmembrane kDa a39 antigen, cell the Then, UltraPure. MicroBeads Anti-Biotin and antibody to block the activation of antigen-presenting cells by T helper cells cells by Thelper cells of antigen-presenting activation the block to labeled magnetically The Separator. of aMACS field magnetic the induced by interaction with CD40 expressed on antigen-presenting on antigen-presenting expressed CD40 with by interaction induced interactions through ligation of CD40. Clone 5C8 has been shown shown been has 5C8 Clone of CD40. ligation through interactions page 1/4

cells.¹ Adding a CD40-blocking antibody during the stimulation stimulation the during antibody aCD40-blocking Adding cells.¹ T cells. MACS Columns and MACS Separators: CD154 Separators: MACS and Columns MACS ▲ phosphate-buffered containing asolution Prepare Buffer: CD154 of Isolation CD154 of activated selection Positive Reagent and instrument requirements instrument and Reagent Background information Background Separation MACS® of the Principle (# (# dextrose formula-A (ACD-A) or citrate phosphate dextrose (CPD). BSA can be be (CPD). can BSA dextrose phosphate (ACD-A) citrate or formula-A dextrose MultiMACS™ Cell24 Separator. Separator. Cell24 MultiMACS™ can also be performed by using the autoMACS Pro or the or the Pro autoMACS the by using performed be also can characterization. use, as air bubbles could block the column. the block could bubbles air as use, replaced by other such as as such proteins other by replaced saline (PBS), pH saline for use. and 2 and be enriched by using MS or LS Columns. Positive selection selection Positive Columns. or LS MS by using enriched be bovine serum (FBS). Buffers or media containing Ca containing media or Buffers (FBS). serum bovine

Note: + 130-091-222). Keep buffer cold (2−8 cold 130-091-222). buffer Keep Solution Rinsing 130-091-376) autoMACS® with 1:20 . Due to its transient expression within hours after activation, activation, after hours within expression transient its to . Due cells are retained within the column. The unlabeled cells cells unlabeled The column. the within retained are cells To increase the purity, the positively selected cell fraction fraction cell selected positively the purity, To the increase

mM EDTA by diluting MACS BSA Stock Solution Solution Stock BSA MACS EDTA by diluting mM EDTA can be replaced by other supplements such as anticoagulant citrate citrate anticoagulant as such supplements other by replaced be can EDTA + + cells can be eluted as the positively selected cell cell selected positively the as eluted be can cells cells are magnetically labeled with CD154-Biotin CD154-Biotin with labeled magnetically are cells 154 + cells is separated over a second column. over asecond separated is cells 7.2, 0.5% bovine serum albumin (BSA), albumin serum 7.2, bovine 0.5% + cells for phenotypical and functional functional and for phenotypical cells human + T cells and plays an important important an plays and Tcells serum albumin, albumin, serum Order no. 130-092-658 + °C). Degas buffer before before buffer °C). Degas 2+ antigen-specific CD4 antigen-specific or Mg or 2+ human are not recommended recommended not are + serum, or fetal fetal or serum, + cells. After After cells. cells can can cells + +

140-001-861.06 ▲ ▲ ▲ When working with tissues or lysed blood, prepare a single-cell asingle-cell prepare blood, or lysed tissues with working When coat, or buffy blood peripheral anticoagulated with working When 2. # 130-092-173). ▲ 2.1.1 2.1.1 2.1 ● ● ● ● ● ● are for research useonlyandnotfor diagnostic ortherapeutic use. For details refer to the protocols section at www.miltenyibiotec. section protocols the to refer For details or the Dead Cell Removal Kit (# Kit 130-090-101). Removal Cell Dead or the density gradient centrifugation, for example, using Ficoll-Paque™. Ficoll-Paque™. using for example, centrifugation, gradient density com/protocols. remove dead cells, we recommend using density gradient centrifugation centrifugation gradient density using we recommend cells, remove dead for example, a sample stimulated with CytoStim (# 130-092-172, 130-092-172, (# CytoStim with stimulated asample for example, stimulus. of the addition for the should be treated exactly the same as the stimulated sample, except except sample, stimulated the as same the exactly treated be should methods. standard using suspension peripheral blood mononuclear cells (PBMCs) should be isolated by (PBMCs) isolated be should cells mononuclear blood peripheral Unless otherwise specificallyindicated,Miltenyi all Unless otherwise Biotec andservices products

Always include a negative control in the experiment. The sample sample The experiment. the in control a negative include Always A positive control should also be included in the experiment, experiment, the in included be also should control A positive Dead cells may bind non-specifically to MACS MicroBeads. To MicroBeads. MACS to non-specifically bind may cells Dead Anti-Biotin-PE (# 130-090-756). For more information For more information 130-090-756). (# Anti-Biotin-PE VarioMACS™ or SuperMACS™ II Separators. For details refer to the respective respective the to refer details For Separators. II SuperMACS™ or VarioMACS™ ▲ 130-094-133) (# grade –functional pure CD40 (Optional) (# 130-092-172, # 130-092-172). CytoStim (Optional) grade PepTivator® –premium pp65 (Optional) CMV 30 Filters, Pre-Separation (Optional) (# Solution Iodide Propidium (Optional) for flow Fluorochrome-conjugated (Optional) Protocol 130-093-438) for stimulation of Tcells. for stimulation # 130-093-438) Sample preparation Sample 2.1.1). 7-AAD for flow cytometric exclusion of dead cells. of dead exclusion cytometric 7-AAD for flow aspirate supernatant. Repeat step. washing ww.miltenyibiotec.com/antibodies. cytometric analysis, e.g., CD4-FITC (# CD4-FITC e.g., analysis, cytometric remove cell clumps. remove cell pellet in buffer and centrifuge at 200×g for 10−15 for 200×g at centrifuge and buffer in pellet antibody to block down-regulation of CD154-expression (see of CD154-expression down-regulation block to antibody to refer antibodies fluorochrome-conjugated about MACS Separator data sheet. data Separator MACS autoMACS Column Multi MS LS Positive selection In vitro Note: Note: ‑ 24

Column adapters are required to insert certain columns into the the into columns certain insert to required are adapters Column To remove after density gradient separation, resuspend cell cell resuspend separation, gradient density after To platelets remove stimulation for induction of CD154 expression for induction stimulation 10⁸ 10⁸ 10⁷ a. number Max. 2×10⁸ of labeledcells 10⁹ Max. number number Max. 4 ×10⁹ 2 ×10⁹ 2 ×10⁸ of total cells µm (#µm 20 at minutes iiAS QuadroMACS, MidiMACS, OctoMACS, MiniMACS, autoMACS Pro, autoMACS VarioMACS, SuperMACS II VarioMACS, SuperMACS II Separator MultiMACS Cell24 MultiMACS 130-080-501) and and 130-080-501) 130-093-233) or 130-041-407) to to 130-041-407) °C. Carefully Carefully °C.

▲ ▲ ▲ ▲ ▲ 1. 1. same the use 10⁷ cells, than fewer with working When cells. 10⁷ total 4. 4. 3. 3.

2. 2. 7. 6. 6. 5. 5. volumes and total volumes). total and volumes numbers, cell higher with working When indicated. as volumes cell labeling. labeling. cell BSA or FBS, because of non-specific stimulation. of non-specific because or FBS, BSA remove cell clumps which may clog the column. Moisten filter with with filter Moisten column. the clog may which clumps remove cell 8. 30 Filters, (Pre-Separation mesh nylon for 2×10⁷ total cells, use twice the volume of all indicated reagent reagent indicated of all volume the twice use for 2×10⁷ cells, total specific cell labeling. Working on ice may require increased increased require may on ice Working labeling. cell specific µm 30 through cells Pass labeling. magnetic before suspension (e.g. accordingly volumes total and volumes reagent up all scale prevent capping of antibodies on the cell surface and non-specific non-specific and surface cell on the of antibodies capping prevent temperatures and/or longer incubation times may lead to non- to lead may times incubation longer and/or temperatures incubation times. times. incubation buffer before use. before buffer page 2/4 page Work fast, keep cells cold, and use pre-cooled solutions. This will will This solutions. pre-cooled use and cold, cells keep Work fast, Do not use media containing any non-human proteins, such as as such proteins, non-human any containing media not use Do For optimal performance it is important to obtain asingle obtain to important it is performance For optimal 2–8 is temperature incubation recommended The Volumes for magnetic labeling given below are for up to are below given labeling Volumes for magnetic 10 minutes. Aspirate supernatant. Aspirate 10 minutes. Add 20 ▲ ▲ Add an antigen or control reagent in the appropriate appropriate the in reagent or control antigen Add an culture in mL per of 10⁷ cells at adensity cells Resuspend µL of buffer per 10⁷ total cells. cells. 10⁷ per total of buffer µL 40 in pellet cell Resuspend for 10 at 300×g suspension cell Centrifuge for 4–16 5% CO₂. at 37 and °C hours cells Incubate grade –functional pure of CD40 µg/mL Add 1 (Optional) Mix well and incubate for 15 incubate and well Mix cells. 10⁷ per total ofAdd CD154-Biotin 10 µL Collect cells carefully by pipetting up and down when working working when down up and by pipetting carefully cells Collect 0.5–1 by adding Wash cells for at 300×g centrifuge and medium by adding Wash cells µL of buffer per 10⁷ total cells. cells. 10⁷ per total of buffer µL 80 in pellet cell Resuspend number. cell Determine (# 130-094-133) to the cell suspension. suspension. (# 130-094-133) cell the to (2−8 antigen-presenting cells. antigen-presenting with smaller volumes. Rinse the dish with cold buffer. Check Check buffer. cold with dish the Rinse volumes. smaller with with CD154 antibodies should be performed immediately after stimulation. stimulation. after immediately performed be should antibodies CD154 with dish again. dish of 5×10⁶ density cells/cm². concentration. cells. cells. completely. for 10 at 300×g centrifuge microscopically for any remaining cells, if necessary, rinse the the rinse necessary, if cells, remaining for any microscopically at a dishes in cells Plate serum. 5% human containing medium regulation of CD154 expression on T cells induced by interaction with CD40 on on CD40 with interaction by induced Tcells on expression CD154 of regulation supernatant completely. supernatant levels are detected 4–16 hours after after 4–16 hours detected are levels Note: Note: °C). °C). CD154 is transiently expressed on activated CD4 activated on expressed transiently is CD154

µL of Anti-Biotin MicroBeads UltraPure per 10⁷ total 10⁷ per total UltraPure MicroBeads of Anti-Biotin µL 2.2 The addition of a CD40-blocking antibody prevents the down- the prevents antibody aCD40-blocking of addition The Magnetic labeling Magnetic mL of buffer per 10⁷ cells and and 10⁷ per cells of buffer mL in vitro in minutes. Aspirate supernatant supernatant Aspirate minutes. minutes in the refrigerator refrigerator the in minutes stimulation. Therefore, staining staining Therefore, stimulation. µm, # µm, minutes. Aspirate Aspirate minutes. 130-041-407) to to 130-041-407) + T cells. The highest highest The Tcells. Order no. 130-092-658 no. Order °C. Higher Higher °C. ‑ cell cell

140-001-861.06 ▲ ▲ 1. 12. 11. 10. 4. 3.

2. 7. 9. 6. 5. are for research useonlyandnotfor diagnostic ortherapeutic use. Refer to the the MultiMACS™ Cell Separator Plus user manual for manual user Plus Separator Cell MultiMACS™ the the to Refer Plus 1.4. section in table the to refer For details cells. Magnetic separation with the MultiMACS™ Cell24 Separator Separator Cell24 MultiMACS™ the with separation Magnetic Columns or LS MS with separation Magnetic according to the number of total cells and the number of CD154 number the and cells of total number the to according to the next step. next the to instructions on how to use the instrument the on how use to instructions Unless otherwise specificallyindicated,Miltenyi all Unless otherwise Biotec andservices products Always wait until the column reservoir is empty before proceeding proceeding before empty is reservoir column the until wait Always Choose an appropriate MACS Column and MACS Separator Separator MACS and Column MACS appropriate an Choose Mix well and incubate for 15 incubate and well Mix ▲ ▲ ▲ Remove column from the separator and place it on a suitable it on asuitable place and separator the from column Remove Collect of buffer. amount appropriate the with Wash column of amount appropriate the with by rinsing column Prepare Apply cell suspension onto the column. Collect flow-through flow-through Collect column. onto the suspension Apply cell Place column in the magnetic field of a suitable MACS MACS of asuitable field magnetic the in column Place To increase purity of CD154 purity To increase (2.3). separation magnetic to Proceed of buffer. µL 500 in up10⁸ to cells Resuspend 1−2 by adding Wash cells Pipette the appropriate amount of buffer onto the column. column. onto the of buffer amount appropriate the Pipette (2−8 Immediately flush out the magnetically labeled cells by firmly by firmly cells labeled magnetically out the flush Immediately data sheet. over a second MS Column. Repeat the magnetic separation separation magnetic the Repeat Column. MS over a second collection tube. 3. step from effluent cells. unlabeled containing completely. for 10 at 300×g centrifuge unlabeled cells that pass through and combine with the the with combine and through pass that cells unlabeled reservoir is empty. is reservoir Separator. For details refer to the respective MACS Column Column MACS respective the to refer For details Separator. procedure as described in steps 1 to 6 by using a new column. anew 6by 1to using steps in described as procedure column. the into plunger the pushing the first onto the second, equilibrated column instead of a collection tube. tube. acollection of instead column equilibrated second, the onto first the buffer:

Note: Note: Note: °C). °C). 2.3

Perform washing steps by adding buffer aliquots only when the column column the when only aliquots buffer adding by steps washing Perform To perform a second column run, you may elute the cells directly from from directly cells the elute may you run, column asecond To perform For higher cell numbers, scale up buffer volume accordingly. accordingly. volume buffer up scale numbers, cell higher For Magnetic separation Magnetic µL 3×500 MS: 5 MS: 1mL MS: µL 00 mL of buffer per 10⁷ cells and and 10⁷ per cells of buffer mL + cells, enrich the eluted fraction fraction eluted the enrich cells, minutes. Aspirate supernatant supernatant Aspirate minutes. minutes in the refrigerator refrigerator the in minutes LS: 3×3 mL LS: 3 mL LS: LS: 5mL LS:

+

▲ ▲ ▲ A) 1. 3. 3.

(# 130-093-438), a CD40 blocking-antibody was added during the the during added was blocking-antibody aCD40 (# 130-093-438), 2. CD154 CD154 MiniMACS™ Separator. Separator. MiniMACS™ were fluorescently stained with Anti-Biotin-PE (# 130-090-756) 130-090-756) (# Anti-Biotin-PE with stained were fluorescently PBMCs were stimulated for 16 hours with PepTivator pp65 for 16 with CMV hours PBMCs were stimulated donor using the CD154 MicroBead Kit, two MS Columns, and a and Columns, MS two Kit, CD154 the MicroBead donor using debris and dead cells were excluded from the analysis based on based analysis the from were excluded cells dead and debris cells. For details refer to the section describing the cell separation separation cell the describing section the to refer For details cells. labeled of magnetically frequency the and labeling, of magnetic Magnetic separation with the autoMACS® Pro Separator Separator Pro autoMACS® the with separation Magnetic scatter signals and propidium iodide fluorescence. fluorescence. iodide propidium and signals scatter of CD154. Subsequently, down-regulation prevent to stimulation and CD4 (VIT4)-APC CD154 CD4 (# 130-092-374)and detect to have a temperature of ≥10 °C. have atemperature programs in the respective user manual. manual. user respective the in programs Separator. Pro autoMACS® the B) page 3/4 page

Program choice depends on the isolation strategy, the strength strength the strategy, isolation on the depends choice Program Buffers used for operating the autoMACS Pro Separator should should Separator Pro autoMACS the for operating used Buffers on how use to for instructions manual user respective the to Refer After separation After Before separation Before Collect positive fraction in row C of the tube rack tube row Cof the in fraction positive Collect Posseld2 selection: Positive For a standard separation choose the following program: following the choose separation For astandard for tubes provide and sample the containing Apply tube Prepare and prime the instrument. the prime and Prepare CD4-APC CD154 the using of aseparation Example

CD4-APC Kit MicroBead collecting the labeled and unlabeled cell fractions. Place sample sample Place fractions. cell unlabeled and labeled the collecting sample pp65–stimulated CMV sample pp65–stimulated CMV tube in row A of the tube rack and the fraction collection tubes tubes collection fraction the and rack tube row Aof the in tube in rows B and C. Band rows in 10 10 10 10 10 10 -1 -1 + + 1 0 1 0 ² ³ ² ³ ¹ ¹ T cells were isolated from human PBMCs of aCMV human from were isolated Tcells cells were separated using the CD154 MicroBead Kit. Cells Cells Kit. CD154 the MicroBead using were separated cells -1 -1 CD154-Biotin/ CD154-Biotin/ 0 0 1 1 10 10 ¹1 ¹1 Anti-Biotin-PE Anti-Biotin-PE 0² 0² 10 10 ³ ³

CD4-APC CD4-APC Unstimulated control Unstimulated control Unstimulated 10 10 10 10 10 10 -1 -1 1 0 1 0 ² ³ ¹ ² ³ ¹ -1 CD154-Biotin/ -1 CD154-Biotin/ 0 0 1 1 Order no. 130-092-658 no. Order 10 10 ¹1 Anti-Biotin-PE ¹1 Anti-Biotin-PE + cells. Cell Cell cells. 0² 0² 10 10 ³ ³ +

140-001-861.06 Reference4. All other trademarks mentioned in this publication are the property of their respective respective their of property the are publication this in mentioned trademarks other All TERMS. BY THESE BOUND BE TO AGREES that warranty no provides Biotec Miltenyi PLACE. IN LICENSES APPROPRIATE Please notice. prior without change to subject are specifications and information All 1. Legal notices Legal customer’s use of this product does not and will not infringe intellectual property property intellectual infringe not will and not does product this of use customer’s the for www.miltenyibiotec.com visit or Support Technical Biotec Miltenyi contact Trademarks Technical information autoMACS, MACS, MidiMACS, MACS, autoMACS, visit or representative Biotec Miltenyi local your contact Please conditions. and terms separate by excluded be may uses Certain limitations. certain have may and/or OF BY apply. USE may terms Additional Warranty”). (“Product order of time the at applicable product’s the of date expiration the on ending and distributor authorized owners and are used for identification purposes only. purposes identification for used are and owners worldwide. countries various in affiliates its and/or Biotec Miltenyi of trademarks or herein. contained omissions or errors editorial are for research useonlyandnotfor diagnostic ortherapeutic use. Miltenyi Biotec provides technical support worldwide. Visit worldwide. support technical provides Biotec Miltenyi www.miltenyibiotec.com/local to find your nearest Miltenyi Biotec Biotec Miltenyi nearest your find to www.miltenyibiotec.com/local Licenses warranty product Limited with Miltenyi Biotec’s published specifications for the product at the time of order, order, of time the at product the for specifications published Biotec’s Miltenyi with (whether the customer is an academic or for-profit entity). This product may not be be not may product This entity). for-profit or academic an is customer the (whether Refer to to Refer The purchase of this product conveys to the customer the non-transferable right to to right non-transferable the customer the to conveys product this of purchase The patents issued or pending more or one by covered be may use its and/or product This Miltenyi by provided statements other and protocols, data, information, technical The APRODUCT IF DETERMINING FOR RESPONSIBLE SOLELY IS CUSTOMER THE TERMS. BY THESE BOUND BE TO AGREES CUSTOMER PRODUCT, THE THIS contact. most up-to-date information on Miltenyi Biotec products. Biotec Miltenyi on information up-to-date most shelf life stated on the product label, packaging or documentation (as applicable) or, (as applicable) documentation or packaging label, product the on stated life shelf rights owned by a third party. BY USE OF THIS PRODUCT, THE CUSTOMER CUSTOMER PRODUCT, THE OFTHIS BY USE party. athird by owned rights use the purchased amount of the product in research conducted by the customer customer the by conducted research in product the of amount purchased the use documentation, applicable its with accordance in conditions and use normal under forth in Miltenyi Biotec’s General Terms and Conditions for the Sale of Products and and Products of Sale the for Conditions and Terms General Biotec’s Miltenyi in forth its or Biotec Miltenyi by product the of delivery of date the on beginning aperiod for substantially conform to and materials and workmanship in defects material from further sold. Additional terms and conditions (including the terms of a Limited Use Use aLimited of terms the (including conditions and terms Additional sold. further Miltenyi Biotec’s website at www.miltenyibiotec.com for more information. more for www.miltenyibiotec.com at website Biotec’s Miltenyi METHODS. set as terms warranty the to subject provided is Warranty Product Biotec’s Miltenyi free be to product this warrant B.V. affiliate(s) Biotec its KG and/or &Co. Miltenyi Copyright © 2020 Miltenyi Biotec and/or its affiliates. All rights reserved. rights All affiliates. its and/or Biotec Miltenyi ©2020 Copyright OctoMACS, QuadroMACS, SuperMACS, and VarioMACS LICENSES ADDITIONAL REQUIRE MAY PRODUCT OFTHIS USE CUSTOMER’S Ficoll-Paque is a trademark of GE Healthcare companies. Healthcare GE of atrademark is Ficoll-Paque RESPONSIBLE FOR DETERMINING FOR ITSELF WHETHER IT HAS ALL ALL HAS IT WHETHER ITSELF FOR DETERMINING FOR RESPONSIBLE APPLICATION AND PURPOSE PARTICULAR CUSTOMER’S FOR SUITABLE IS Label License) may apply. may License) Label information such of completeness or accuracy the but reliable, be to believes Biotec Miltenyi which experience or tests, information, on based are document this in Biotec Unless otherwise specificallyindicated,Miltenyi all Unless otherwise Biotec andservices products judgment to the information. Miltenyi Biotec shall not be liable for any technical or or technical any for liable be not shall Biotec Miltenyi information. the to judgment DEPENDING ON THE SPECIFIC APPLICATION. THE CUSTOMER IS SOLELY SOLELY IS CUSTOMER THE APPLICATION. SPECIFIC THE ON DEPENDING is not guaranteed. Such technical information and data are intended for persons with with persons for intended are data and information technical Such guaranteed. not is Warranty”). (“Product delivery of date from (1) ONE YEAR thereof, absence the in Services available on Miltenyi Biotec’s website at www.miltenyibiotec.com, as in effect effect in as www.miltenyibiotec.com, at website Biotec’s Miltenyi on available Services knowledge and technical skills sufficient to assess and apply their own informed informed own their apply and assess to sufficient skills technical and knowledge Frentsch, M. M. Frentsch, according to CD154 expression. Nat Med. 11: 1118–1124. Med. Nat expression. CD154 to according antigens www.miltenyibiotec.com et al. et (2005) Direct access to CD4 to access Direct (2005) the Miltenyi Biotec logo, Biotec Miltenyi the for all data sheets and protocols. protocols. and sheets data for all

+ T cells specific for defined defined for specific Tcells MiniMACS, MultiMACS,

are registered trademarks trademarks registered are

page 4/4 page Order no. 130-092-658 no. Order