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. 2022 Feb 17;3(1):101163. doi: 10.1016/j.xpro.2022.101163
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-Human CD45 (HI30)-89Y—100 Tests (1:100) Fluidigm Cat#3089003B
Anti-Human CD196/CCR6 (G034E3)-141Pr—50 Tests (1:100) Fluidigm Cat#3141003A
Anti-Human CD19 (HIB19)-142Nd—100 Tests (1:100) Fluidigm Cat#3142001B
Anti-Human CD45RA (HI100)-143Nd—100 Tests (1:100) Fluidigm Cat#3143006B
Anti-Human CD69 (FN50)-144Nd—100 Tests (1:100) Fluidigm Cat#3144018B
Anti-Human CD4 (RPA-T4)-145Nd—100 Tests (1:100) Fluidigm Cat#3145001B
Anti-Human IgD (IA6-2)-146Nd—100 Tests (1:100) Fluidigm Cat#3146005B
Anti-Human CD20 (2H7)-147Sm—100 Tests (1:100) Fluidigm Cat#3147001B
Anti-Human CD14 (RMO52)-148Nd—100 Tests (1:100) Fluidigm Cat#3148010B
Anti-Human CD56/NCAM (NCAM16.2)-149Sm—100 Tests (1:100) Fluidigm Cat#3149021B
Anti-Human CD138/Syndecan-1 (DL-101)-150Nd—25 tests (1:100) Fluidigm Cat#3150012C
Anti-Human TNFa (mab11)-152Sm—100 Tests (1:100) Fluidigm Cat#3152002B
Anti-Human TIGIT (MBSA43)-153Eu—100 Tests (1:100) Fluidigm Cat#3153019B
Anti-Human CD3 (UCHT1)-154Sm—100 Tests (1:100) Fluidigm Cat#3154003B
Anti-Human CD27 (L128)-155Gd—100 Tests (1:100) Fluidigm Cat#3155001B
Anti-Human IFNg (B27)-158Gd—100 Tests (1:100) Fluidigm Cat#3158017B
Anti-Human CD161 (HP-3G10)-159Tb—100 Tests (1:100) Fluidigm Cat#3159004B
Anti-Human/Mouse MIP1beta (D21-1351)-160Gd—100 Tests (1:100) Fluidigm Cat#3160013B
Anti-Human/Mouse Tbet (4B10)-161Dy—100 Tests (1:100) Fluidigm Cat#3161014B
Anti-Human FoxP3 (259D/C7)-162Dy—50 Tests (1:100) Fluidigm Cat#3162024A
Anti-Human CD183/CXCR3 (G025H7)-163Dy—100 Tests (1:100) Fluidigm Cat#3163004B
Anti-Human IL-17A (N49-653)-164Dy—100 Tests (1:100) Fluidigm Cat#3164002B
Anti-Human CD279/PD-1 (EH12.2H7)-165Ho—100 Tests (1:100) Fluidigm Cat#3165042B
Anti-Human CD197/CCR7 (G043H7)-167Er—50 Tests (1:100) Fluidigm Cat#3167009A
Anti-Human CD8 (SK1)-168Er—100 Tests (1:100) Fluidigm Cat#3168002B
Anti-Human CD25 (2A3)-169Tm—100 Tests (1:100) Fluidigm Cat#3169003B
Anti-Human Granzyme B (GB11)-171Yb—100 Tests (1:100) Fluidigm Cat#3171002B
Anti-Human CD57 (HCD57)-172Yb—100 Tests (1:200) Fluidigm Cat#3172009B
Anti-Human HLA-DR (L243)-173Yb—100 Tests (1:200) Fluidigm Cat#3173005B
Anti-Human CD94 (HP-3D9)-174Yb—100 Tests (1:100) Fluidigm Cat#3174015B
Anti-Human Perforin (B-D48)-175Lu—100 Tests (1:100) Fluidigm Cat#3175004B
Anti-Human CD127/IL-7Ra (A019D5)-176Yb—100 Tests (1:100) Fluidigm Cat#3176004B
Anti-Human CD16 (3G8)-209Bi—100 Tests (1:100) Fluidigm Cat#3209002B
Anti-Granzyme K antibody [GM-24C3] (ab3771) (1:100) Abcam Cat#ab3771
Chemicals, peptides, and recombinant proteins
Ficoll-Paque Plus Global Life Sciences Cat#45001740
1× Phosphate-Buffered Saline without Calcium & Magnesium Mediatech Cat #21040CV
10× RBC Lysis Buffer BioLegend Cat #420301
0.4% Trypan Blue Solution Life Technologies Cat #15250061
autoMACS Running Buffer – MACS Separation Buffer Miltenyi Biotec Cat #130091221
CD138 MicroBeads, Human Miltenyi Biotec Cat #130051301
DMSO Sigma-Aldrich Cat #D2650
PBS, pH 7.4 (1×) Gibco Cat#10010023
Bovine Serum Albumin (BSA), 30% ± 2% in 0.85% NaCl Sigma-Aldrich Cat#A7284
Sodium Azide, 10% (w/v) solution in Ultra-Pure H2O Teknova Cat#S0209
RPMI 1640 (1×) Medium Gibco Cat#11875093
Fetal Bovine Serum, heat inactivated, qualified (FBS) Gibco Cat#10438026
Antibiotic-Antimycotic (100×) Gibco Cat#15240096
UltraPureTM DNAse/RNAse-Free Distilled Water Invitrogen Cat#10977023
ViaStainTM AOPI Staining Solution Nexcelom Bioscience Cat#CS2-0106-5mL
Cell-IDTM Intercalator-103Rh—500 μM Fluidigm Cat#201103A
Cell-IDTM Intercalator-191/193Ir—125 μM Fluidigm Cat#201192A
Maxpar® Cell Acquisition Solution Fluidigm Cat#201237
Maxpar® Cell Staining Buffer Fluidigm Cat#201068
EQTM Four Element Calibration Beads Fluidigm Cat#201078
eBioscienceTM Permeabilization Buffer (10×) Invitrogen Cat#00833356
eBioscienceTM Fixation/Permeabilization Concentrate Invitrogen Cat#00512343
eBioscienceTM Fixation/Permeabilization Diluent Invitrogen Cat#00522356
Human TruStain FcXTM (FcR Blocking Solution) BioLegend Cat#422302
PierceTM 16% Formaldehyde Solution (w/v), Methanol-free Thermo Scientific Cat#28906
Critical commercial assays
Maxpar® X8 Antibody Labeling Kit, 166Er—4 Rxn Fluidigm Cat#201166A
Biological samples
Human bone marrow samples (sex, age as required par study) Any supplier N/A
Software and algorithms
FlowJo Becton Dickinson & Company https://www.flowjo.com/solutions/flowjo/downloads/
CyTOF Software v7.0.8493 Fluidigm https://www.fluidigm.com/software
Other
SepMate-50 Tubes Stemcell Technologies Cat #85460
10 mL Serological Pipet Corning Inc. Cat #357530
25 mL Serological Pipet Corning Inc. Cat #1367530
25 mL Serological Pipet Corning Inc. Cat #1367530
MACS SmartStrainer 100 μm Miltenyi Biotec Cat #130098463
autoMACS Pro Miltenyi Biotec Cat #130092545
CoolCell FTS30 Thermo Fisher ScientificTM Cat #07210008
Countess II Automated Cell Counter Thermo Fisher ScientificTM Cat #AMQAX1000
Countess Cell Counting Chamber Slides Thermo Fisher ScientificTM Cat #C10228
Falcon 96-Well Polystyrene Microplate, TC Treated Corning Inc. Cat #353296
15 mL Polypropylene Centrifuge Tubes Corning Inc. Cat#430052
50 mL Polypropylene Centrifuge Tubes Corning Inc. Cat#430829
Cellometer® SD100 Cell Counting Chamber Slides Nexcelom Bioscience CHT4-SD100-014
70 μm Cell Strainer, Polypropylene Frame Biologix Cat#15-1070
Eppendorf® 1.5 mL Safe-Lock Microcentrifuge Tubes, natural Eppendorf Cat#022363204
96-well V-Bottom Plate, untreated Corning Inc. Cat#3896
Polystyrene 96-well Microplate Corner Notch Lid Corning Inc. Cat#3930
Falcon® 5 mL Round Bottom Polypropylene Tubes (FACS tubes) Corning Inc. Cat#352063
Falcon® 5 mL Round Bottom Polystyrene Tubes with 35 μm Cell Strainer Snap Caps Corning Inc. Cat#352235
ThermoFisher ScientificTM SorvallTM LegendTM XTR Refrigerated Centrifuge, 120VAC Thermo Fisher ScientificTM Cat#75004521
FisherbrandTM Mini-Centrifuge 100–240V, 50/6-Hz Universal Plug, Grey Thermo Fisher ScientificTM Cat#12-006-901
Fisher Scientific Digital Vortex Mixer Thermo Fisher ScientificTM Cat#0215370
Thermo ScientificTM PrecisionTM Circulating Water Bath Thermo Fisher ScientificTM Cat#TSCIR19
Cellometer® Auto 2000 Cell Viability Counter Nexcelom Bioscience Auto 2000
Fluidigm HeliosTM Mass Cytometer Fluidigm N/A
Nebulizer for HeliosTM Mass Cytometer Fluidigm Cat#107144
WB Injector for HeliosTM Mass Cytometer Fluidigm Cat#107950