ver.1.4 The path to new discoveries From basic to clinical research in immunology Good research starts with happy cells.
gentlemacs TM Octo Dissociator with Heaters TM Whole Blood MicroBeads 37 RNA Option 0.25mL 90-95% automacs Pro Separator Whole Blood 40 Tissue Storage Solution #130-100-008 Dissociation Kit MACS gentlemacs Dissociator Lamina Propria Dissociation Kit & gentlemacs Dissociator P4P8WEB MACSxpress Isolation Kit 20 30 ml 8 ml 5 min (15 min) MACSxpress Separator PI CD45 + cells Live cells Live cells SSC Figure 1 C57BL/6Lamina Propria Dissociation Kit (gentlemacs Dissociator Propidium Iodide Solution (130-093-233) MACSQuant Analyzer 2.010 7 80 %viability FSC TCRβ-APC-Cy7 CD45R(B220)-VioBlue 130-100-008 MACS Tissue Storage Solution 130-105-808 Adipose Tissue Dissociation Kit () 130-103-464 Epidermis Dissociation Kit () 130-095-928 Epidermis Dissociation Kit () 130-097-410 Lamina Propria Dissociation Kit () 130-105-807 Liver Dissociation Kit () 130-095-927 Lung Dissociation Kit () 130-095-926 Spleen Dissociation Kit () 130-095-929 Tumor Dissociation Kit () 130-096-730 Tumor Dissociation Kit () 130-101-540 Whole Skin Dissociation Kit () CD11c-VioBlue CD11b-APC-Vio770 CD15-PE MACSxpress Neutrophil Isolation Kit A B C (%) (%) 100 10.5 8.5 95 6.5 4.5 90 2.5 0.5 85 0.2 80 0.1 75 CD16-APC 70 0.0 Figure 2 (A) (CD15 + CD16 + )MACSQuant Analyzer(B) 6 982%)9523%)981%)1 ml2.1-3.310 6 (C) 130-104-434 MACSxpress Neutrophil () 130-101-319 MACSxpress Starting Kit MACSxpress 2
2MACS25 MACS MACS Technology MACS Technology A B C S N S N 50 nm a b MACS Technology 1 50 nm) Figure 4 MACS (A) MACS (B) (C) (a) (b)20 MACS Technology Truly Untouched Cells S N S N Figure 3 PBMCCD14/CD16(red) (green)amacs B MACS Technology viability automacs MS Column LS Column LD Column Deplete Deplete_S Deplete_05 Deplete_025 1 Target 10 6 cells in 10 8 cells Flow Sorting Total 206 40 MACS & Flow Sorting Total 47 166 2 automacs Pro 5 & 40 1 ~10 5 cells 10 9 10 10 MiniMACS Separator automacs Pro Separator Buffy coatleucopac MultiMACS Cell24 Separator Plus MultiMACS X 3
MACS at a Glance & NK cells Human PBMCs MACSprep PBMC # 130-115-169 CD4 + CD4 # 130-045-101 Whole Blood CD4 # 130-090-877 CD4 + T Cell # 130-096-533 MACSxpress CD4 T Cell # 130-098-195 Treg cells CD3 # 130-050-101 Whole Blood CD3 # 130-090-874 CD4 + CD25 + Regulatory T Cell # 130-091-301 CD4 + CD25 + CD127 dim/ Regulatory T Cell # 130-094-775 CD25 + CD49d Regulatory T Cell # 130-094-551 CD4 + CD25 + CD45RA + Regulatory T Cell # 130-093-631 MACSxpress Treg # 130-109-557 CD4 + T cell subsets Naive Naive Pan T Cell # 130-097-095 REAlease CD3 # 130-117-038 CD8 + CD2 # 130-091-114 Pan T Cell # 130-096-535 CD8 # 130-045-201 Whole Blood CD8 # 130-090-878 CD8 + T Cell # 130-096-495 MACSxpress CD8 T Cell # 130-098-194 γ/δ + Anti-TCRγ/δ # 130-050-701 TCRγ/δ + T Cell # 130-092-892 DN Double-negative T Cell # 130-092-614 CD8 + T cell subsets CD6 # 130-091-264 MACSxpress Pan T Cell # 130-098-193 NK CD3 + CD56 + NKT Cell # 130-093-064 Anti-iNKT # 130-094-842 ILC2 ILC2 # 130-114-825 NK cells CD56 # 130-050-401 Whole Blood CD56 # 130-090-875 NK Cell # 130-092-657 MACSxpress NK Cell # 130-098-185 NK cell subsets CD56 + CD16 NK Cell # 130-092-661 CD56 + CD16 + NK Cell # 130-092-660 CD56 + CD8 + /CD8 NK Cell # 130-092-659 CD56 + CD57 + NK Cell #130-093-395 CD57 #130-092-073 CD4 + RTE CD4 + Recent Thymic Emigrant # 130-094-299 Naive CD4 + Naive CD4 + T Cell # 130-094-131 Skin-homing CD4 + CLA + T Cell # 130-092-435 Anti-CLA # 130-092-464 Naive CD8 + Naive CD8 + T Cell # 130-093-244 Memory CD8 + CD8 + Memory T Cell # 130-094-412 Other Memory CD4 + Memory CD4 + T Cell # 130-091-893 MACSxpress CD4 Memory T Cell # 130-106-770 TEM cells CD4 + Effector Memory T Cell # 130-094-125 TCM cells CD4 + Central Memory T Cell # 130-094-302 TEMRA cells CD8 + CD45RA + Effector T Cell # 130-094-485 Cytotoxic CD8 + CD57 + T Cell # 130-093-396 NK : NK Cell #130-092-657 REAlease CD56 #130-117-033 TH1 cells IFN-γ Secretion Assay # 130-054-201 TH2 cells CD294 (CRTH2) # 130-091-274 IL-4 Secretion Assay # 130-054-101 TH17 cells IL-17 Secretion Assay # 130-094-542 Effector CD27 # 130-051-601 Activated T cell subsets Other CD25 # 130-092-983 CD30 # 130-051-401 CD69 # 130-092-355 CD154 # 130-092-658 CD137 # 130-093-476 CD8 + T CD8 + T Cell # 130-096-495 REAlease CD8 #130-117-036 CD4 + T CD4 + T Cell # 130-096-533 REAlease CD4 #130-117-037 CD28 #130-093-247 CD62L # 130-091-758 CD45RA # 130-045-901 CD45RO # 130-046-001 CD127 #130-094-945 4
Mouse NK cells CD3ε # 130-094-973 Pan T Cell # 130-095-130 CD90.1 # 130-094-523 CD90.2 # 130-049-101 CD5 (Ly-1) # 130-049-301 CD4/CD8 (TIL) # 130-116-480 CD49b (DX5) # 130-052-501 NK Cell # 130-115-818 Anti-NKp46 # 130-095-390 CD4 + CD4 (L3T4) # 130-117-043 CD4 + T Cell # 130-104-454 CD4 (TIL) # 130-116-475 Treg cells CD4 + CD25 + Regulatory T Cell # 130-091-041 CD25 # 130-091-072 CD8 + CD8a (Ly-2) # 130-117-044 CD8a + T Cell # 130-104-075 CD8 (TIL) # 130-116-478 γ/δ + TCRγ/δ + T Cell # 130-092-125 ink NK1.1 + inkt Cell # 130-096-513 T cell subsets MACS () Naive CD4 + Naive CD4 + T Cell #130-104-453 CD4 + CD62L + T Cell # 130-106-643 Memory : CD4 + or CD8 + T Cell CD62L (L-selectin) # 130-049-701 Naive CD8 + Naive CD8a + T Cell #130-096-543 Activated CD25 # 130-091-072 CD154 # 130-093-129 untouched 2 TH1 cells Mouse IFN-γ Secretion Assay # 130-090-517 TH17 cells Mouse IL-17 Secretion Assay # 130-094-213 TH2 cells Mouse IL-4 Secretion Assay # 130-090-515 MACSxpress MACSxpressMACS MACSxpress2 MACS Cytokine Secretion Assays : + CD45Catch Reagent37 C Catch Reagent Anti-PE 5
& NK cells PBMC Pan T, CD4 + T, CD8 + T, NK (Untouched) 18 (PBMC) CD56 () (CD56 + ) CD4 + T Cell () (PBMC) (CD3 + CD4 + ) min fast kit! CD56-PE CD56-PE CD3-FITC CD3-FITC CD3-FITC CD3-FITC Figure 7 CD56 + NK, NKT CD4-PE CD4-PE Figure 5 CD4 + T CD4 + T Cell () () (CD3ε + CD4 + ) CD4-APC CD4-APC CD3ε-PE CD3ε-PE min fast kit! Research diversity: & & T 2 CD45RA-FITC Naive CD4 + T Cell (PBMC) CD45RA-FITC (CD4 + CD45RA + ) min fast kit! Figure 6 CD4 + T MACSQuant Analyzer BALB/cC57BL/6 CD3ε + CD4 + 95% CD11cAnti-MHC Anti-TCRγ/δ CD4-APC CD4-APC Figure 8 CD4 + T >96%CD4 + MACSTNK Naive CD4 + T Cell () (CD4 + CD62L + CD44 -/low ) min fast kit! RA T T T NK In vitro polarization Th T GvHD GvHD HIV HIV CD3ε-APC-Vio770 CD4-VioBlue CD3ε-APC-Vio770 CD4-VioBlue Figure 9 CD4 + T MACSQuant AnalyzerBALB/cC57BL/6CD3ε + CD4 + CD62L + 95%CD4 + CD25 + regulatory T(<0.3% CD44-FITC CD62L-PE 6
T cell subsets of your choice T Progressive regulatory T cell research: T T (Tregs) FoxP3 expansion CD4 + CD25 + Regulatory T Cell Isolation Kit ( ) CD4 + CD25 + Regulatory T Cell Isolation Kit ( ) PBMCCD4 + TCD25 + / CD25 - Tregs Responder TCD4 + CD25 - Tregs CD4 + CD25 + CD127 dim/- Regulatory T Cell Isolation Kit ( ) CD127 dim/- FoxP3Tregs In vitro expansion CD4-FITC CD4 + CD25 + CD127 dim/- T CD25-APC Figure 10 T MACSxpress Treg Isolation Kit ( ) T MACSxpress30 MACSxpress Treg CD25-APC gated on CD4 + cells CD127-PE Handling time (min) Anti-FoxP3-APC gated on CD4 + cells CD127-PE Competitor Kit A CD4 + CD25 + Regulatory T Cell PBMC Competitor Kit B 0 30 60 90 120 150 180 MHC multimer 2 CD154-APC (PBMC) Anti-IL-17-PE IL-17 CD154-APC Anti-IL-17-PE Figure 12 IL-17 PBMCCytoStim (#130-092-172) CD40CD40 IL-17 CD40 (#130-094-133) CD154CD4 + TCD154 + IL-17 + 0.113%90% TH1 (IFN-γ), TH2 (IL-4), TH17 (IL-17), IL-10 secreting Tr1 cytotoxic T TH17TH1 EAE Tr1 Tr1 Tr1 T gated on CD4 + cells MACST MHC multimer-peanti-pe (#130-048-801) CD154 CD154 () (#130-092-658) CD154 () (#130-093-129) Aspergillus-reactive CD4 + T CD154-APC Anti-IL-17-PE T Treg detection Kit (PE/APC)( ) Treg detection Kit (PE/APC)( ) CD4 CD154 CD154 FoxP3, CD4, CD25FcR CD154 IL-10 IFN-γ CD25-PE Treg Detection Kit (APC) () Anti-FoxP3-APC gated on CD4 + cells Figure 13 AspergillusCD4 + T PBMCAspergillus lysate6 CD4 + T CD154CD4 + CD154 + 0.35% CD154 () (#130-092-658) MS CD154 + CD4 + CD154 + 29.2% Anti-IL-10 (#130-096-042) Anti-IFN-γ (#130-113-493) CD4-FITC CD25-PE Figure 11 PBMCT 7
MACS at a Glance & NK cells Human Hematopoietic stem cells CD133 Hematopoietic Tissue # 130-100-830 Diamond CD133 # 130-094-913 CD34 # 130-046-702 (Large Scale # 130-046-703) CD34 UltraPure # 130-100-453 CD34 MultiSort Kit # 130-056-701 CD34 + CD38 - Cell # 130-114-822 Diamond CD34 # 130-094-531 Lineage Cell # 130-092-211 Leukocytes CD45 # 130-045-801 Whole Blood CD45 # 130-090-872 Tumor-Infiltrating Leukocytes CD45 (TIL) # 130-118-780 Myeloid progenitor cells CD33 # 130-045-501 Whole Blood CD33 # 130-094-773 Antigen-presenting cells Anti-HLA-DR # 130-046-101 Erythroblasts CD71 # 130-046-201 Erythrocytes CD235a (Glycophorin A) # 130-050-501 Megakaryocytes CD61 # 130-051-101 Thrombocytes CD61 # 130-051-101 Anti-CX3CR1 # 130-099-032 Other CD123 # 130-094-432 Granulocytes CD15 # 130-046-601 Whole Blood CD15 # 130-091-058 Basophils Basophil # 130-092-662 Diamond Basophil # 130-094-434 Eosinophils CD16 # 130-045-701 Eosinophil # 130-092-010 MACSxpress Eosinophil # 130-104-446 Neutrophils CD16 # 130-045-701 CD66abce # 130-092-393 Whole Blood CD66b # 130-104-913 MACSxpress Neutrophil # 130-104-434 CD10 # 130-093-452 Monocytes/ Macrophages CD14 # 130-050-201 CD14 UltraPure # 130-118-906 Pan Monocyte # 130-096-537 Whole Blood CD14 # 130-090-879 CD11b # 130-049-601 Classical Monocyte # 130-117-337 CD16 + Monocyte # 130-091-765 Slan (M-DC8) + Monocyte # 130-093-026 Myeloid dendritic cells Myeloid Dendritic Cell # 130-094-487 MDC subsets MDC1s CD1c (BDCA-1) + Dendritic Cell # 130-090-506 # 130-119-475 MDC2s CD141 (BDCA-3) # 130-090-512 Dendritic cells Blood Dendritic Cell # 130-091-379 Pan-DC # 130-100-777 Plasmacytoid dendritic cells Plasmacytoid Dendritic Cell # 130-097-415 Diamond Plasmacytoid Dendritic Cell # 130-097-240 CD304 (BDCA-4/ Neuropilin-1) # 130-090-532 CD303 (BDCA-2) # 130-090-509 Tissue dendritic cells CD209 (DC-SIGN) # 130-092-868 CD1a # 130-051-001 CD207 (Langerin) # 130-097-898 CD19 # 130-050-301 Whole Blood CD19 # 130-090-880 REAlease CD19 # 130-117-034 CD20 # 130-091-104 CD22 # 130-046-401 Pan B Cell # 130-101-638 B Cell # 130-091-151 B-CLL Cell # 130-103-466 MACSxpress Pan B Cell # 130-104-444 MACSxpress B Cell # 130-098-190 MACSxpress B-CLL Cell # 130-104-445 Naive/ resting Naive B Cell # 130-091-150 CD43 # 130-091-333 Anti-IgD # 130-103-775 MACSxpress Naïve B Cell # 130-098-186 Activated B cell subsets B Pan B Cell # 130-101-638 CD25 # 130-092-983 CD30 # 130-051-401 CD69 # 130-092-355 Memory Memory B Cell # 130-093-546 IgM + Memory B Cell # 130-093-619 IgG + Memory B Cell # 130-094-350 Switched Memory B Cell # 130-093-617 CD27 # 130-051-601 Anti-IgG # 130-047-501 Anti-IgM # 130-093-230 Plasma cells CD138 # 130-051-301 Whole Blood & Bone Marrow CD138 # 130-105-961 CD38 # 130-092-263 Plasma Cell # 130-093-628 8
Mouse Hematopoietic stem cells Anti-Sca-1 (FITC) # 130-092-529 CD105 MultiSort Kit (PE) # 130-092-924 CD117 # 130-091-224 Lineage Cell # 130-090-858 Direct Lineage Cell # 130-110-470 Leukocytes CD45 # 130-052-301 Tumor-Infiltrating Leukocytes CD45 (TIL) # 130-110-618 Myeloid cells Antigen-presenting cells Anti-MHC Class # 130-052-401 B cell Progenitors CD93 # 130-105-753 Erythrocytes Anti-Ter-119 # 130-049-901 Megakaryocytes CD61 # 130-109-678 Myeloid-derived suppressor cells Myeloid-Derived Suppressor Cell # 130-094-538 Neutrophils Anti-Ly-6G # 130-092-332 Anti-Ly-6B.2 (7/4) # 130-100-781 Neutrophil # 130-097-658 Monocytes/ Macrophages Anti-F4/80 UltraPure # 130-110-443 Macrophage (Peritoneum) # 130-110-434 CD11b # 130-049-601 CD115 # 130-096-354 Monocyte (BM) # 130-100-629 Dendritic cells CD11c UltraPure # 130-108-338 Pan DC # 130-092-465 Pan Dendritic Cell # 130-100-875 CD45R (B220) # 130-049-501 CD19 # 130-052-201 CD22 # 130-103-776 Pan B Cell # 130-104-443 Other CD23 # 130-098-784 Anti-Siglec-F # 130-118-513 Indirect Conventional dendritic cells CD8 + DCs Anti-XCR1 (Spleen) # 130-115-721 CD8 + Dendritic Cell # 130-091-169 Plasmacytoid dendritic cells Anti-mPDCA-1 # 130-091-965 Plasmacytoid Dendritic Cell # 130-107-093 B-2 cells B-1a cells CD5 (Ly-1) # 130-049 301 B-1a Cell # 130-097-413 B-2 cell subsets 1 Anti- ( web ) Anti-FITC MultiSort Kit # 130-058-701 Anti-PE MultiSort Kit # 130-090-757 Anti-APC MultiSort Kit # 130-091-255 Anti-Biotin MultiSort Kit # 130-091-256 Anti-Biotin # 130-090-485 Anti-Biotin UltraPure # 130-105-637 Anti-FITC # 130-048-701 Anti-PE # 130-048-801 Anti-PE UltraPure # 130-105-639 Anti-APC # 130-090-855 Anti-Cy5/Anti-Alexa Fluor 647 # 130-091-395 Anti-Cy7 # 130-091-652 CD4 + DCs CD4 + Dendritic Cell # 130-091-262 MACS () untouched 2 MACSxpress Tissue dendritic cells Epidermal Langerhans Cell # 130-095-408 CD207 (Langerin) # 130-097-572 MACSxpressMACS MACSxpress2 Resting B Cell # 130-090-862 CD43 (Ly-48) # 130-049-801 GC Germinal Center B Cell (PNA) # 130-110-479 Memory Memory B Cell # 130-095-838 Anti-Mouse IgG1 # 130-047-101 Anti-Mouse IgG2a+b # 130-047-201 Plasma cells CD138 + Plasma Cell # 130-092-530 CD138 # 130-098-257 Regulatory Regulatory B Cell # 130-095-873 MZ and FO MZ and FO B Cell # 130-100-366 9
DC & Monocytes / Macrophages untouchedpositive (DC) PBMC DC PBMC32 pan-monocyte Pan-DC Pan Monocyte (PBMC) (Pan DC) (PBMC) (Pan monocyte) CD303 (BDCA-2)-FITC CD1c (BDCA-1)-PE CD303 (BDCA-2)-FITC CD1c (BDCA-1)-PE CD303 (BDCA-2)-FITC CD141 (BDCA-3)-APC CD14-FITC CD16-PE CD14-FITC CD16-PE CD14-FITC Anti-Slan (M-DC8)-APC Figure 14 pan DC CD1c + CD20 MDC1CD141 + MDC2CD303 + PDC3CD14 + (88.6%, n=13) Figure 17 (Pan-monocyte) Classical (CD14 ++ CD16 ), non classical (CD14 + CD16 ++ ), intermediate (CD14 ++ CD16 + ) monocytes Pan Dendritic Cell (Pan DC) () Anti-F4/80 UltraPure () () (F4/80 + ) CD11c-APC CD11c-APC CD11b-APC CD11b-APC Anti-mPDCA-1-FITC Anti-mPDCA-1-FITC Anti-F4/80-FITC Anti-F4/80-FITC Figure 15 pan DC CD11c hi conventional DCmPDCA-1 + CD11c low Plasmacytoid DC Myeloid Dendritic Cell Plasmacytoid Dendritic Cell MDCPDC 2p.8-9 MACS T (DC) Blood Dendritic Cell Enumeration Kit ( 3DC Figure 18 TAM) F4/80UltraPure MACS () in vitro Mo-DC CytoBox Mo-DC GM-CSFIL-4 Mo-DC Differentiation Inspector ( ) Mo-DC GM-CSF+IL-4 TNF-α or LPS CD1c (BDCA-1)-PE MDC1s PDCs CD303 (BDCA-2)-FITC CD141 (BDCA-3)-APC MDC2s CD303 (BDCA-2)-FITC DC PDC CD303 (BDCA-2) MDC1 CD1c (BDCA-1) MDC2 CD141 (BDCA-3) MFI Mo-DC Mo-DC CD14 CD209 CD83 381,6 1,4 0,5 MFI 804,2 301,1 7,8 370,5 3,2 2,9 Monocytes immo-dcs mmo-dcs Monocytes immo-dcs mmo-dcs Monocytes immo-dcs mmo-dcs MFI Figure 16DC Figure 19 10
& & BPlasma BPan Bresting B B Pan B Cell (PBMC) (B&) Eosinophil (PBMC) () CD19-APC CD19-APC CD43-PE CD43-PE Figure 20Pan-B BCD43 + BPlasma (>95%, n=5) MZ and FO B Cell () (MZ B) (FO B) Figure 22Eosinophil () Myeloid-Derived Suppressor Cell () (Gr-1 high Ly-6G + ) (Gr-1 dim Ly-6G ) CD21/CD35-PE-Cy7 CD21/CD35-PE-Cy7 CD21/CD35-PE-Cy7 Anti-Gr-1-PE Anti-Gr-1-PE Anti-Gr-1-PE Side scatter Side scatter CD16-FITC CD16-FITC CD23-PE CD23-PE CD23-PE CD11b-APC CD11b-APC CD11b-APC Figure 21BB CD21 hi CD23 lo/ marginal zone (MZ) BCD21 int CD23 hi follicular (FO) B 12 Figure 23Myeloid-Derived Suppressor Cell (MDSC) MDSC2 12 371 MACS Marker Screen () 1 MACS Marker Screen371 96 well plate 1 MACSQuant all-in-one solution 4 MACS Marker Screen Plate 1 A B C D E F G H 1 2 3 4 5 6 7 8 9 10 11 12 Empty CD1a CD1b CD1c (BDCA-1) CD1d CD2 CD3 CD4 (VIT4) CD5 CD6 CD7 CD8 CD9 CD10 CD11a CD11b CD11c CD13 CD14 CD15 CD16 CD18 CD19 CD20 CD21 CD22 CD23 CD24 CD25 CD26 CD27 CD28 CD29 CD30 CD31 CD32 CD33 CD34 CD35 CD36 CD37 CD38 CD39 CD40 CD41a CD41b CD42a CD42b CD43 CD44 CD45 CD45RA CD45RB CD45RO CD46 CD47 CD48 CD49a CD49b CD49c CD49d CD49e CD49f CD51 CD51/ CD61 CD52 CD53 CD54 (ICAM-1) CD61 CD62E CD62L CD62P CD63 CD64 CD66acde CD66b CD66c CD68 CD69 CD70 CD71 CD72 CD73 CD74 CD79a CD79b CD80 CD81 CD82 CD83 CD84 CD55 (DAF) CD56 CD57 CD58 (LFA-3) CD85a (ILT5) doclogic gatelogic TM TM platelogic TM graphlogic TM 11
Research Translational research Clinical MACS Sample Preparation Tissue dissociation Automated cell processing CliniMACS Prodigy MACS Cell Separation Cell isolation or depletion Cell isolation & phenotyping MACSQuant Flow Sorting MACS Cell Culture MACS Flow Cytometry T / TLR 3 Vio Dyes REA 1 Culture or expansion Phenotyping of cells GMP compliant tools GCTP/MF IL-4/GM-CSF/TNF-α Texpansion kit CD3 T / MACS GMP Cytokine MACS GMP Cell Culture & Stimulation MACSmolecular KIR Molecular applications Cryopreservation CryoMACS DMSO 10 (EP, USP) CryoMACS Freezing Products Viscover Imaging Agents CT, MRI,, Imaging MACS GMPGMP GMP 135-0041 16-10 NEX5F TEL 03-5646-8910FAX 03-5646-8911 www.miltenyibiotec.com E-mail macs@miltenyibiotec.jp 03-5646-9606 0120-03-5645 03-5646-8566 AM900PM500 201806dE200