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Prussian blue iron staining kit

Manufactured by Solarbio
Sourced in China

The Prussian blue iron staining kit is a laboratory product designed to detect and visualize the presence of iron in biological samples. It provides a simple and reliable method for identifying iron through the formation of a characteristic blue pigment. The kit includes all the necessary reagents and materials to perform the staining procedure.

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6 protocols using prussian blue iron staining kit

1

Cytotoxicity Evaluation of MIONs@PDA in HUMSCs

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Briefly, HUMSCs were obtained after normal deliveries following 38–40 week gestations. HUMSCs were cultured in Dulbecco’s modified Eagle medium (Gibco, Waltham, MA, USA) containing 10% fetal bovine serum (Gibco) and negative for mycoplasma contamination. All experiments were conducted using HUMSCs at passages 5–8. After reaching 80% confluency, various concentrations of MIONs@PDA (0, 10, 25, 50, 100, and 200 μg/mL) were added to the medium for 12 h. The Cell Counting Kit-8 (Sigma-Aldrich, St. Louis, MO, USA) assay was used to detect the cytotoxicity of MIONs@PDA. HUMSCs were stained with the Prussian blue iron staining kit (Solarbio, Beijing, China), according to the manufacturer’s instructions. The phenotype of HUMSCs was confirmed by the expression of surface markers (positive for CD44-fluorescein isothiocyanate (FITC) and CD105-phycoerythrin (PE), and negative for CD45-FITC) (all from BD Biosciences, San Jose, CA, USA) using a FACSC anto II flow cytometer (FC500; Beckman Coulter Brea, CA, USA), and the data were analyzed with the CXP software (Beckman Coulter). Differentiation capacity of the HUMSCs was assessed by inducing osteocyte and adipocyte differentiation using the StemPro Osteogenesis/Adipogenesis Differentiation Kit (Invitrogen, Waltham, MA, USA) and evaluated by Alizarin Red S, Oil red O, and Alcian blue staining, respectively.
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2

Spinal Cord Injury Histopathology Assessment

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The animals were anesthetized and transcardially perfused with 4% PFA. 8-mm segments of spinal cord at the center of injury site (or the same position in sham tissues) were collected and sectioned (30 µm thick). Immunohistochemistry of 4-Hydroxynonenal (4-HNE) and CTSB was conducted using the Super Plus™ High Sensitive and Rapid Immunohistochemical Kit (pH6.0) (Elabscience Biotechnology, E-IR-R221). The anti-4 HNE antibody and anti-CTSB antibody were used for labelling (Supplementary Table 2). Secondary antibody-only controls were employed to validate antibody specificity and distinguish genuine target staining from the background. Histopathological tissue staining was conducted using the Prussian Blue Iron staining kit (Solarbio Science & Technology, G1422) and Modified Oil Red O staining kit (Solarbio Science & Technology, G1261) following the manufacturer's instructions. The positively stained areas in the injured epicenter were quantitatively analyzed using the ImageJ software "IHC profiler."
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3

Prussian Blue Iron Staining of MSCs

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MSCs grown to 80% confluency were incubated with Fe3O4 NPs (50 µg/mL) for 16 h, washed three times with PBS, and stained with a Prussian blue iron staining kit (Solarbio, Beijing, China), according to the manufacturer’s instructions.
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4

Cellular Iron Deposition Analysis

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According to the instructions, Prussian Blue Iron Staining Kit (Solarbio, Beijing, China) was used to determine cellular iron deposition. The cells were fixed with 4% paraformaldehyde (PFA) for 20 minutes, and washed twice with tap water and twice with deionized water. Then the cells were incubated with Perls stain for 30 minutes at room temperature.
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5

Evaluating Fe3O4 Nanoparticle Uptake and Toxicity in MSCs

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Adherent MSCs (80–90% confluence) were washed with phosphate-buffered saline (PBS) and incubated with Fe3O4 NPs (0, 25, 50, 100, 150, and 200 µg/mL) in FBS-free culture medium for 2 h (n = 4). The cells and NPs were then co-cultured in a fresh medium containing 10% FBS for 14 h. The MSCs were washed with PBS and stained using the Prussian blue iron staining kit (Beijing Solarbio Science and Technology, Co. Ltd., Beijing, China), according to the manufacturer’s protocol.
In vitro toxicity experiments were performed at 24 h after Fe3O4 NP (0, 25, 50, 100, 150, and 200 µg/mL) labeling. Cytotoxicity of Fe3O4 NPs was evaluated using the cell counting kit (CCK)-8 assay (KeyGEN BioTECH, Nanjing, China). Fe3O4 NPs that displayed minimal cytotoxicity were used in the subsequent experiments.
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6

Staining MSCs with Prussian Blue

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MSCs grown to 80 % confluency were incubated with Fe 3 O 4 NPs (50 µg/mL) for 16 h as described previously, [47] washed three times with phosphate-buffered saline (PBS), and stained with a Prussian blue iron staining kit (Solarbio, Beijing, China), according to the manufacturer's instructions.
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