1 mL Luer-Lok™ syringe sterile, single use, polycarbonate |
BD |
309628 |
|
5 mL Luer-Lok™ syringe sterile, single use, polycarbonate |
BD |
309646 |
|
Alexa Fluor 488 C5 maleimide |
Invitrogen |
A10254 |
For synthesis of fluorescently-labeled tetrazine |
Alexa Fluor 647 Phalloidin |
Invitrogen |
A22287 |
For staining cell culture samples |
Aluminum foil |
VWR |
89107–726 |
|
Biopsy punch with plunger, 1.0 mm |
Integra Miltex |
69031–01 |
|
Biopsy punch, 4 mm |
Integra Miltex |
33–34 |
|
Blunt needle, 23G 0.5”, Non-Sterile, Capped |
SAI Infusion Technologies |
B23-50 |
|
Bottle-top vacuum filter, 0.22 μm |
Corning |
CLS430521 |
|
Calcium chloride |
VWR |
1B1110 |
For microgel washing buffer |
Capillary-piston assemblies for positive-displacement pipettes, 1000 μL max. volume |
Rainin |
17008609 |
|
Capillary-piston assemblies for positive-displacement pipettes, 25 μL max. volume |
Rainin |
17008605 |
|
Capillary-piston assemblies for positive-displacement pipettes, 250 μL max. volume |
Rainin |
17008608 |
|
Centrifuge tube, 15 mL |
CELLTREAT |
667015B |
|
Centrifuge tube, 50 mL |
CELLTREAT |
229421 |
|
Chloroform, ACS grade, Glass Bottle |
Stellar Scientific |
CP-C7304 |
For synthesis of fluorescently-labeled tetrazine |
Corona plasma gun, BD-10A High Frequency Generator |
ETP |
11011 |
|
CryoTube Vials, Polypropylene, Internal Thread with Screw Cap |
Nunc |
368632 |
|
D1 mouse mesenchymal cells |
ATCC |
CRL-12424 |
Example cell line for culture in MAP gels |
DAPI |
Sigma-Aldrich |
D9542 |
For staining cell culture samples |
Deuterium oxide, 99.9 atom % D |
Sigma-Aldrich |
151882 |
For NMR spectroscopy |
Dialysis tubing, regenerated cellulose membrane, 12-14 kDa molecular weight cut-off |
Spectra/Por |
132703 |
For purifying HA-NB and HA-Tet |
Diethyl ether |
VWR |
BDH1121-4LPC |
For synthesis of fluorescently-labeled tetrazine |
Dimethylformamide |
Sigma-Aldrich |
277056 |
For synthesis of fluorescently-labeled tetrazine |
4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM) |
TCI-Chemicals |
D2919 |
For modifying HA |
Dithiothreitol (DTT) |
Thermo Scientific |
R0861 |
Non-degradable dithiol linker (substitute for MMP-cleavable peptide) |
Dulbecco’s Modified Eagle’s Medium - high glucose, w/ 4500 mg/L glucose, L-glutamine, sodium pyruvate, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture |
Sigma-Aldrich |
D6429–500ML |
For D1 cell culture |
EMS Paraformaldehyde, Granular |
VWR |
100504–162 |
For making 4% PFA |
Ethanol absolute (200 proof) |
KOPTEC |
89234–850 |
|
Heating Plate |
Kopf Instruments |
HP-4M |
|
2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) |
Sigma-Aldrich |
H3375 |
|
Sodium hyaluronate, 79 kDa average molecular weight, produced in bacteria Streptococcus zooepidemicus, pharmaceutical grade, microbial contamination <100 CFU/g, bacterial endotoxins <0.050 IU/mg |
Contipro |
N/A |
79 kDa average molecular weight was used for HA-Tet synthesis, but these methods but could be adapted for other molecular weights. |
IMARIS Essentials software package |
Oxford Instruments |
N/A |
Microscopy image analysis software |
Infusion pump, dual syringe |
Chemyx |
N/A |
|
Kimwipe |
Kimberly-Clark |
34120 |
|
Laboratory stand with support lab clamp |
Geyer |
212100 |
|
Liquid nitrogen |
Airgas |
NI 180LT22 |
|
Lithium Phenyl(2,4,6-trimethylbenzoyl)phosphinate |
TCI-Chemicals |
L0290 |
|
Methyltetrazine-PEG4-maleimide |
Kerafast |
FCC210 |
For synthesis of fluorescently-labeled tetrazine |
2-(4-Morpholino)ethane Sulfonic Acid (MES) |
Fisher Scientific |
BP300–100 |
For modifying HA |
Micro cover glass, 24×60mm No. 1 |
VWR |
48393–106 |
|
Mineral oil, heavy |
Sigma-Aldrich |
330760 |
|
MMP-cleavable dithiol crosslinker peptide (Ac-GCRDGPQGIWGQDRCG-NH2) |
GenScript |
N/A |
|
5-Norbornene-2-methylamine |
TCI-Chemicals |
95–10–3 |
For HA-NB synthesis |
Packing tape |
Scotch |
3M 1426 |
|
Parafilm |
Bemis |
PM996 |
|
PEG(thiol)2 |
JenKem Technology USA |
A4001-1 |
For synthesis of fluorescently-labeled tetrazine |
Petri dish, polystyrene, disposable, Dia. x H=150 × 15 mm |
Corning |
351058 |
|
Pluronic F-127 |
Sigma-Aldrich |
P2443 |
For washing HMPs |
Phosphate buffered saline (PBS) 1x |
Gibco |
10010023 |
|
RainX water repellent glass treatment |
Grainger |
465D20 |
|
RGD peptide (Ac-RGDSPGERCG-NH2) |
GenScript |
N/A |
|
Rubber bands |
Staples |
112417 |
|
Sodium chloride |
Chem-Impex |
30070 |
For dialysis |
Span® 80 for synthesis |
Sigma-Aldrich |
1338–43–8 |
|
Sylgard 184 Silicone Elastomer |
Electron Microscopy Science |
4019862 |
|
Syringe filter, Whatman™ Uniflo, 0.2 μm PES, 13 mm diameter |
Cytvia |
09–928–066 |
|
Tetraview LCD digital microscope |
Celestron |
44347 |
|
Tetrazine-amine HCl salt |
Chem-Impex |
35098 |
For HA-Tet synthesis |
Triethylamine |
Sigma-Aldrich |
471283 |
For synthesis of fluorescently-labeled tetrazine |
Tris(2-carboxyethyl)phosphine (TCEP) |
Millipore Sigma |
51805–45–9 |
|
Triton X-100 |
VWR |
97063–864 |
|
Trypan blue solution, 0.4% |
Thermo Fisher Scientific |
15250061 |
|
Trypsin EDTA (0.25%), Phenol red |
Fisher Scientific |
25–200–056 |
For lifting adherent cells to seed in MAP gels |
Tygon® formula E-3603 laboratory tubing |
Millipore Sigma |
Z765244 |
|
UV curing system controller, LX500 LED |
OmniCure |
010–00369R |
|
UV curing head, LED spot UV |
OmniCure |
N/A |
|
UV light meter, Traceable |
VWR |
61161–386 |
|
Vacuum dessicator |
Bel-Art |
08–594–15C |
|
X-Acto Z Series Precision Utility Knife |
Elmer’s |
XZ3601W |
|