Human collagen type I (BD Bioscience, San Jose, CA) was diluted to the appropriate concentration using sterile 4 mM HCl and 10× RPMI was added to neutralize pH. Freshly trypsinized cells re-suspended in FBS were added to the neutralized collagen I. The collagen gel was then allowed to polymerize at 37°C, 5% CO2, 90% humidity overnight. To harvest the cells from collagen gels, collagenase (Sigma-Aldrich) was added and incubated at 37°C for 15 min. Cell pellet was harvested from gel by centrifuge at 300g for 5 min.
Human collagen type 1
Manufactured by BD
Sourced in Germany
Human collagen type I is a natural component of the human extracellular matrix. It is a structural protein that provides strength and support to various tissues in the human body.
Automatically generated - may contain errors
Lab products found in correlation
Collagenase, by Merck Group (1 mentions)
Incb28060, by Selleck Chemicals (1 mentions)
Anti erk1 2 antibody, by Santa Cruz Biotechnology (1 mentions)
Human epidermal growth factor (hegf), by Merck Group (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Insulin, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
β glycerol phosphate, by Merck Group (1 mentions)
Hepatocyte growth factor (hgf), by R&D Systems (1 mentions)
2 protocols using human collagen type 1
1
Collagen-Based 3D Cell Culture and Harvest
2
Cell Culture and Inhibitor Protocols
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The cell culture media RPMI 1640 and DMEM/F12 were purchased from Gibco, Life Technologies (Grand Island, NY); α-Modified Essential Medium (α-MEM) from Sigma Aldrich (Schnelldorf, Germany). Penicillin/streptomycin was obtained from Gibco, Life Technologies (Grand Island, NY); FBS from PAA Laboratories (Cölbe, Germany). Other media supplements (including insulin, hydrocortisone, cholera toxin, hEGF, β -glycerol phosphate, ascorbic acid, and dexamethasone) were purchased from Sigma Aldrich (Schnelldorf, Germany). Human collagen type I was obtained from BD Biosciences (Heidelberg, Germany).
INCB28060 was purchased from Selleck Chemicals (Munich, Germany) and prepared as a 5 mM stock solution in 100% dimethyl sulfoxide (DMSO) and stored at—80°C [34 ]. INCB28060 was used at a concentration of 100 nM unless otherwise specified. HGF was purchased from R&D Systems (Minneapolis, MN), diluted in phosphate-buffered saline (PBS) with 1% bovine serum albumin (BSA) and stored at -20°C per manufacturer´s instructions.
Antibodies against human phosphorylated MET and total MET were obtained from Cell Signaling Technology (Boston, MA, USA). Anti-ERK 1/2 antibody was purchased from Santa Cruz Biotechnology (Heidelberg, Germany).
INCB28060 was purchased from Selleck Chemicals (Munich, Germany) and prepared as a 5 mM stock solution in 100% dimethyl sulfoxide (DMSO) and stored at—80°C [34 ]. INCB28060 was used at a concentration of 100 nM unless otherwise specified. HGF was purchased from R&D Systems (Minneapolis, MN), diluted in phosphate-buffered saline (PBS) with 1% bovine serum albumin (BSA) and stored at -20°C per manufacturer´s instructions.
Antibodies against human phosphorylated MET and total MET were obtained from Cell Signaling Technology (Boston, MA, USA). Anti-ERK 1/2 antibody was purchased from Santa Cruz Biotechnology (Heidelberg, Germany).
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