To evaluate the affinity of FGF2-SPION to hPSCs, cells were seeded into 24 well plate at a seeding density of 4,000 cells/well and starved for 24 h. hPSCs were then incubated with 5 ng/ml TGF-β for 24 h and then they were washed three times with PBS. Next, hPSCs were incubated with SPION or FGF2-SPION in DPBS (Lonza) with 2 mM Mn2+ (Sigma) and 2 mM Ca2+ (Sigma) at RT for 1 h or 2 h. As controls, non- TGF-β treated hPSCs were incubated with SPION or FGF2-SPION at RT for 1 h. Cells were then washed thrice with DPBS and fixed with 4% formaldehyde in DPBS for 15 mins. After fixation, cells were washed with thrice with DPBS. Prussian blue iron staining kit containing potassium ferocyanide and hydrochloric acid in 1:1 ratio (Sigma Aldrich) was applied in each well and let at RT for 30 mins. Finally, the cells were washed with demi water and mounted with Aquatex mounting medium (Sigma Aldrich). Images were made the next day with Nikon E400 microscope (Nikon, Tokyo, Japan). Quantitative analyses were performed using ImageJ software (NIH, Bethesda, MD).
Prussian blue iron staining kit
Manufactured by Merck Group
The Prussian blue iron staining kit is a laboratory reagent used to detect and visualize the presence of iron in biological samples. It provides a reliable and standardized method for staining iron deposits, allowing for the identification and quantification of iron content in tissues or cells.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco phosphate buffered saline (dpbs), by Merck Group (1 mentions)
E400 microscope, by Nikon (1 mentions)
Aquatex mounting medium, by Merck Group (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Lonza (1 mentions)
Astra 6, by Wyatt Technology (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Heleos 2, by Wyatt Technology (1 mentions)
Optilab t rex, by Wyatt Technology (1 mentions)
2 protocols using prussian blue iron staining kit
1
Quantifying FGF2-SPION Affinity in hPSCs
2
Molecular Weight Analysis of Dextran Derivatives
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The number average molecular weight (Mn) and the molar-mass dispersity values of dextran derivatives 5 and 7 were measured by using a SEC column (TSKgel G2500PW and G6000PW columns) coupled with a differential refractometer (RI, Optilab T-rEX, Wyatt Technology) with a laser at λ = 658 nm, thermostated at 25°C, and a multi-angle laser light scattering instrument (MALLS, HELEOS II, Wyatt Technology) equipped with a laser operating at λ = 664 nm. Degassed and filtered (0.1 μm membrane) 0.15 M ammonium acetate/0.20 M acetic acid buffer (pH = 4.5) was used as eluent at a flow rate of 0.5 mL/min (with a refractive index increment value, dn/dc of 0.147 ml/g for dextran).These eluents were also used as solvent of samples, and the resulting solutions were filtered on a 0.45 μm membrane before injection. Finally, 200 μL of each sample at 1 mg/mL were injected. The data have been exploited thanks to the ASTRA 6.1.7.17 software (Wyatt Technology).
Cell culture. Murine macrophage cell line J774A.1 (ATCC) were grown in Dulbecco's Modified Eagle's Medium (Thermo Fischer) supplemented with 10% v/v decomplemented fetal bovine serum (FBS) at 37 °C in humidified conditions with 5% CO2. Prussian blue iron staining kit (Sigma-Aldrich) containing potassium ferrocyanide, pararosaniline and hydrochloric acid was used for cell staining..
Cell culture. Murine macrophage cell line J774A.1 (ATCC) were grown in Dulbecco's Modified Eagle's Medium (Thermo Fischer) supplemented with 10% v/v decomplemented fetal bovine serum (FBS) at 37 °C in humidified conditions with 5% CO2. Prussian blue iron staining kit (Sigma-Aldrich) containing potassium ferrocyanide, pararosaniline and hydrochloric acid was used for cell staining..
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