The protein levels were determined by ELISA according to the manufacturer’s
instructions for the respective ELISA Kit, summarized inTable 3 . The protein levels in a
sample are determined by comparing it to a serially diluted standard solution
with a defined protein concentration by optical density using a plate reader
(Infinite M Plex, TECAN, Männedorf, Switzerland). For this, cell-seeded
scaffolds were prepared as described previously and incubated in the presence or
absence of fibrin for 1 and 7 days.
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instructions for the respective ELISA Kit, summarized in
sample are determined by comparing it to a serially diluted standard solution
with a defined protein concentration by optical density using a plate reader
(Infinite M Plex, TECAN, Männedorf, Switzerland). For this, cell-seeded
scaffolds were prepared as described previously and incubated in the presence or
absence of fibrin for 1 and 7 days.
Summarized proteins and ELISA Kits.
| Protein | Full name | Kit | Manufacturer | Range | Sensitivity |
|---|---|---|---|---|---|
| HF1a | Hypoxia-·lnduced factor 1-alpha | Human HIFIA ELiSA kit | Invitrogen, thermo Fisher Scientific Waltham, MA, USA | 81.92-20.000 pg/mL | <30 pg/mL |
| PROX1 | Prospero homobox 1 | PROX1 ELISA kit | ELAab Science lnc. Housto, Texas, USA | 0 156-10 ng/mL | <0.057 ng/mL |
| VEGFA | Vascular endothelial growth factor A | Human VEGF-A cell Lysates ELISA kit | Invitrogen, thermo Fisher Scientific Waltham, MA, USA | 8.23-6000 pg/mL | <10 pg/mL |
| VEGFB | Vascular endothelial growth factor B | Human VEGF-B ELISA kit | Invitrogen, thermo Fisher Scientific Waltham, MA, USA | 0.4-100 ng/mL | <0.4 ng/mL |
| VEGFC | Vascular endothelial growth factor C | Human VEGF-C ELISA kit | Invitrogen, thermo Fisher Scientific Waltham, MA, USA | 0.23-15.0 ng/mL | <0.057 ng/mL |
| FLTI (VEGFR) | FMS related receptor tyrosine kinase 1 | Human VEGFR1/Fit-1 quantikine ELISA kit | R&D Systems Minneapolis, MN, USA | 31.3-2.000 pg/mL | <8.46 pg/mL |
| KDR (VEGFR2) | Kinase insert domain receptor | Human VEGFR2/QRD quantikine ELISA kit | R&D Systems Minneapolis, MN, USA | 78.1-5000 pg/mL | <11.4 pg/mL |
| FLT4 (VEGFR3) | FMS related kinase 4 (FLT4) | Human sVEGFR3/Fit-4 Duoset ELISA kit | R&D Systems Minneapolis, MN, USA | 0.9 pg/mL-50ng/mL | <90 pg/mL |
