REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Biotin mouse lineage panel, diluted 1:40∗ | BD Biosciences | Cat# 559971 RRID: AB_10053179 |
PE streptavidin; diluted 1:100 | BD Biosciences | Cat# 554061 RRID: 554061 |
PE-Cy7 rat anti-mouse Ly-6A/E (Sca1) (clone D7); diluted 1:40∗ | BD Biosciences | Cat# 558162; RRID:AB_647253 |
APC anti-mouse CD117 (c-kit) (clone ACK2); diluted 1:40∗ | BioLegend | Cat# 135108; RRID:AB_2028407 |
Pacific Blue anti-mouse CD150 (clone TC15-12F12.2); diluted 1:40∗ | BioLegend | Cat# 115924; RRID:AB_2270307 |
APC-Cy7 anti-mouse CD48 (clone HM48-1); diluted 1:40∗ | Thermo Fisher Scientific | Cat# 47-0481-82; RRID:AB_2573962 |
Rabbit anti-human/mouse phospho-histone H2A.X (Ser139) (20E3); diluted 1:1000 | Cell Signaling Technology | Cat# 9718S; RRID:AB_2118009 |
Donkey anti-Rabbit IgG, Alexa Fluor 488; diluted 1:500 | Thermo Fisher Scientific | Cat# A21206; RRID:AB_2535792 |
Biological samples | ||
Human whole bone marrow and blood from healthy donors | Lonza | Cat# 1M-105 |
Chemicals, peptides, and recombinant proteins | ||
FLT3-ligand, human | PeproTech | Cat# 300-19 |
IL-6, human | PeproTech | Cat# 200-06 |
SCF, human | PeproTech | Cat# 300-07 |
TPO, human | PeproTech | Cat# 300-18 |
SCF, mouse∗ | PeproTech | Cat# 250-03-10UG |
TPO, mouse∗ | PeproTech | Cat# 315-14-10UG |
Mitomycin C | Sigma-Aldrich | Cat# M0503 CAS:50-07-7 |
Triton X-100 | Sigma-Aldrich | Cat# T8787 CAS:9002-93-1 |
Sodium hydroxide (NaOH) pellets | VWR Life Science | Cat#0583-500G CAS: 1310-73-2 |
Bovine serum albumin (BSA) | Sigma-Aldrich | Cat# A9647 CAS: 9048-46-8 |
Ammonium chloride solution | STEMCELL Technologies | Cat# 07800 CAS: 12125-02-9 |
EDTA (0.5 M) pH 8 | Boston BioProducts | Cat# BM150 CAS: 6381-92-6 |
Methanol | Fisher Scientific | Cat# A412-4 CAS: 67-56-1 |
70% Ethanol | Fisher Scientific | Cat# BP82011 CAS: 64-17-5 |
16% Paraformaldehyde (formaldehyde) aqueous solution | Electron Microscopy Sciences | Cat# 15710 CAS: 50-00-0 |
Experimental models: organisms/strains | ||
Fancd2-/- and WT mice. C57BL/6 background, 2–6-month-old. Any gender. | Parmar et al., 2009 | N/A |
Software and algorithms | ||
FlowJo, version 10.5.3 | N/A | https://www.flowjo.com/ |
OpenComet plugin | N/A | http://www.cometbio.org/ |
ImageJ | N/A | http://fiji.sc |
Other | ||
EasySep Kit, human | STEMCELL Technologies | Cat# 19356 |
EasySep Buffer | STEMCELL Technologies | Cat# 20144 |
EasySep Magnet | STEMCELL Technologies | Cat# 18000 |
MidiMACS Separator | Miltenyi | Cat# 130-042-302 |
MACS MultiStand | Miltenyi | Cat# 130-042-303 |
CD34 MicroBead Kit, human | Miltenyi | Cat# 130-046-702 |
FcR Blocking Reagent, human | Miltenyi | Cat# 130-059-901 |
Lineage Cell Depletion Kit, mouse∗ | Miltenyi | Cat# 130-090-858 |
LS Column Miltenyi | Miltenyi | Cat# 130-042-401 |
MS Column Miltenyi | Miltenyi | Cat# 130-042-201 |
StemSpan SFEM II | STEMCELL Technologies | Cat# 09655 |
MethoCult H4434 Classic | STEMCELL Technologies | Cat# 04434 |
Methocult GF M3434 methylcellulose∗ | STEMCELL Technologies | Cat# 03444 |
PBS pH 7.4 | Gibco | Cat# 100-10-023 |
SYBR-Green dye | Invitrogen | Cat# S7567 |
Fetal bovine serum (FBS) | Sigma-Aldrich | Cat# F2442 |
Hanks balanced salt solution (HBSS) | Lonza | Cat# 10-547F |
HEPES Buffer | Fisher Scientific | Cat# BP299-100 |
Penicillin-streptomycin (PenStrep) | Gibco | Cat# 15140-122 |
L-Glutamine | Gibco | Cat# 25030081 |
70 μm Cell strainer | Fisher Scientific | Cat# 087712 |
10 mL Eccentric tip syringe | BD | Cat# 305482 |
18G 1 Inch needle | BD | Cat# 305195 |
500 mL Bottle Top Vacuum Filter, 0.45 μm | Corning | Cat# 430514 |
CometAssay Single Cell Gel Electrophoresis Assay Kit | Trevigen | Cat# 4250-050-K |
Normal goat serum (10%) | Life Technologies | Cat# 50062Z |
SuperFrost Plus Slides | Fisher Scientific | Cat# 12-550-15 |
ProLong Gold Antifade Mountant with DAPI | Life Technologies | Cat# P36941 |
Double Cytofunnel. Disposable sample chambers with white filter cards | Thermo Scientific | Cat# 5991039 |
Round-Bottom Polystyrene Test Tubes with Cell Strainer Snap Cap, 5mL | Fisher Scientific | Cat# 08-771-23 |
SmartDish | STEMCELL Technologies | Cat# 27370 |
Non-tissue culture treated plate, 12 well flat bottom | Falcon | Cat# 351143 |
STEMvision | STEMCELL Technologies | Cat# 22000 |
Cytospin 3 | Shandon Scientific | Cat# 74000102 |
∗ Indicates reagents exclusive for mouse experiments