Potassium ferrocyanide

Manufactured by Thermo Fisher Scientific

Sourced in United States

Potassium ferrocyanide is a chemical compound with the formula K4[Fe(CN)6]. It is a crystalline, yellow-to-orange salt that is soluble in water. Potassium ferrocyanide is commonly used as a laboratory reagent and in various industrial applications.

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18 protocols using potassium ferrocyanide

Quantifying Cellular Senescence via SA-β-Galactosidase Staining

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Cellular senescence was quantified using cytochemical senescence‐associated β‐galactosidase staining. After microparticle treatment, HUVECs were washed twice with PBS and incubated with 2 mL of fixative (2% formaldehyde and 0.2% glutaraldehyde) for 5 minutes. Fixed cells were washed twice with PBS and then incubated for 14 hours with 2 mL of freshly prepared staining solution (1 mg/mL 5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐galactopyranoside in dimethylformamide, 40 mmol/L citric acid/sodium phosphate, 5 mmol/L potassium ferrocyanide, 5 mmol/L potassium ferricyanide, 150 mmol/L NaCl, and 2 mmol/L MgCl₂) (ThermoFisher). The staining solution was then removed, and cells were washed twice with PBS and once with methanol and allowed to air dry. Cells were visualized by light microscopy (Zeiss, Thornwood, NY) and quantified in 5 random image fields containing a minimum of 100 cells per field for each condition. All images were quantified by a blinded investigator (K.A.S). Cells with blue cytoplasmic staining were identified as senescent‐positive cells. Senescent cells (percentage) were determined as senescence‐associated β‐galactosidase–positive cells/the total number of cells counted.24

Hijmans J.G., Stockelman K.A., Garcia V., Levy M.V., Brewster L.M., Bammert T.D., Greiner J.J., Stauffer B.L., Connick E, & DeSouza C.A. (2019). Circulating Microparticles Are Elevated in Treated HIV‐1 Infection and Are Deleterious to Endothelial Cell Function. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 8(4), e011134.

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Prussian Blue Staining of Tissue

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Tissue sections were submerged in a 1:1 ratio of 20% hydrochloric acid (Thermo Fisher Scientific) and 10% potassium ferrocyanide (Thermo Fisher Scientific) solution for 20 minutes at room temperature. Slides were washed with water, dehydrated with ethanol, and cleared with xylene before being coverslipped with Permount (Thermo Fisher Scientific).

Cerdeño V.M., Hong T., Amina S., Lechpammer M., Ariza J., Tassone F., Noctor S.C., Hagerman P, & Hagerman R. (2018). Microglial Cell Activation and Senescence are Characteristic of the Pathology FXTAS. Movement disorders : official journal of the Movement Disorder Society, 33(12), 1887-1894.

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Synthesis of Inorganic Nanomaterials

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All chemicals were used as purchased without further purification. Chromium potassium sulfate dodecahydrate, sodium chloride (99%), potassium hydrogen carbonate (99.7%), potassium ferrocyanide (99%), bismuth powder (99.5%, 100 mesh), copper powder (99.5%, 100 mesh), tin powder (99.5%, 100 mesh), acetone (99%), and 1-octadecene (90% technical grade) were purchased from Thermo Fisher Scientific (Alfa Aesar, Acros Organics). 1,3-Propanediamine-N,N,N′,N′-tetraacetic acid (99%), 1-octanethiol (98.5%), oleylamine (70% technical grade), bismuth neodecanoate, bismuth nitrate pentahydrate (99.99%), ethylene glycol (99%), and molybdenum carbide were purchased from Merck (Sigma Aldrich). Isopropanol (99.5%) was purchased from Honeywell. Potassium hydroxide (reagent grade) was purchased from VWR. Gold mesh (99.9%, 0.06 mm diameter, 20 × 20 mm area, 82 wires per inch) and bismuth foil (99.999%, 0.25 × 10 × 10 mm) were purchased from Goodfellow Cambridge Ltd.

Potter M., Smith D.E., Armstrong C.G, & Toghill K.E. (2023). Electrochemically decoupled reduction of CO2 to formate over a dispersed heterogeneous bismuth catalyst enabled via redox mediators. Ees Catalysis, 2(1), 379-388.

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Quantifying Cellular Senescence with SA-β-Gal Staining

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Cellular senescence was quantified using cytochemical senescence‐associated β‐galactacidase (SA‐β‐gal) staining (Dimri et al. 1995; Debacq‐Chainiaux et al. 2009). Briefly, subconfluent cells were washed twice with 2 mL of PBS followed by a 5‐min incubation in 2 mL of 2% formaldehyde and 0.2% glutaraldehyde to fix cells. Fixed cells were washed twice with 2 mL of PBS and then incubated for 14 h with 2 mL of freshly prepared staining solution (1 mg/mL 5‐bromo‐4‐chloro‐3‐indolyl‐βD‐galactopyranoside in dimethylformamide, 40 mmol/L citric acid/sodium phosphate, 5 mmol/L potassium ferrocyanide, 5 mmol/L potassium ferricyanide, and 150 mmol/L NaCl, 2 mmol/L MgCl₂) (ThermoFisher, Waltham MA). The staining solution was then removed and cells were washed twice with 2 mL of PBS and once with 1 mL of methanol and allowed to air dry. Cells were visualized by light microscopy (Zeiss, Thornwood, NY) and quantified in five random image fields for each condition. Cells with blue cytoplasmic staining were identified as senescent positive cells. Senescent cells (%) were determined as SA‐β‐gal positive cells divided by the total number of cells counted (Dimri et al. 1995).

Hijmans J.G., Stockleman K., Reiakvam W., Levy M.V., Brewster L.M., Bammert T.D., Greiner J.J., Connick E, & DeSouza C.A. (2018). Effects of HIV‐1 gp120 and tat on endothelial cell sensescence and senescence‐associated microRNAs. Physiological Reports, 6(6), e13647.

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Histochemical Analysis of Brain Tissue

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The rats were perfused with 4% paraformaldehyde (MilliporeSigma), and the brain was removed, fixed with 4% paraformaldehyde for 6 hours, and dehydrated by immersion in 20% sucrose and then in 30% sucrose. The whole brain tissue was sliced coronally into 30 μm thick sections on a cryotome (Thermo Fisher Scientific). The sections were fixed with 4% paraformaldehyde, washed with double distilled water for 30 seconds, and incubated in freshly prepared solutions of 2% hydrogen chloride and 2% potassium ferrocyanide (Thermo Fisher Scientific) for 30 minutes. The hydrogen chloride and potassium ferrocyanide solutions were omitted when preparing the negative control sections. The 20-minute elimination of endogenous peroxidase activity was performed with 99% methanol and 1% hydrogen peroxide (Wang et al., 2015). The diaminobenzidine reaction products were observed under a fluorescence microscope (DM4000 B, Leica), and images were captured at a final magnification of 200×. The data were collected from three fields of view per rat.

Hu C.B., Jiang H., Yang Y., Wang G.H., Ji Q.H., Jia Z.Z., Shen L.H, & Luo Q.Q. (2022). DL-3-n-butylphthalide alleviates motor disturbance by suppressing ferroptosis in a rat model of Parkinson’s disease. Neural Regeneration Research, 18(1), 194-199.

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Synthesis and Characterization of Redox-Active Hydrogel

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All chemicals were used without additional purification. Carboxy-PEG 12-Thiol, methanol (ACS Reagent), and ethanol (ACS Reagent) were purchased from ThermoFisher Scientific. Trans-resveratrol (99%) was purchased from Sigma Aldrich. α-cyclodextrin (98%) was purchased from Tokyo Chemical Industry Co. Both Potassium ferrocyanide (ACS Reagent) and Potassium ferricyanide (98%) were purchased from Acros Organics. Phosphate Buffered Saline Solution (PBS) was prepared using a standard protocol from chemicals purchased from ThermoFisher Scientific (pH 7.4 and conductivity 14 mS/cm). Raw, unprocessed pooled human urine from 12 donors was provided by Lee BioSolutions (pH 6.3 and conductivity 12.12 mS/cm).

Panahi Z., Merrill M.A, & Halpern J.M. (2020). Reusable Cyclodextrin-Based Electrochemical Platform for Detection of trans-Resveratrol. ACS applied polymer materials, 2(11), 5086-5093.

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Cell Viability Assessment using Alamar Assay

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Cell viability was assessed using a 10X Alamar reagent: 650 μM rezasurin (Sigma Aldrich 199303), 78 μM methylene blue (Sigma Aldrich M9140), 1 mM potassium ferrocyanide (ACROS Organics 196781000), and 1 mM potassium ferricyanide (ACROS Organics 424130050) in DPBS, diluted to a final concentration of 1X in the cell culture medium (US patent no. 5501959). After addition of the Alamar reagent, cells were incubated in a humidified incubator with 5% CO2 at 37°C for 1 to 6 h before reading the fluorescence of the solution using the Promega GloMax Discover Microplate Reader (Ex: 525 nm; Em: 580–640 nm).

Davis A., Morris K.V, & Shevchenko G. (2022). Hypoxia-directed tumor targeting of CRISPR-Cas9 and HSV-TK suicide gene therapy using lipid nanoparticles. Molecular Therapy. Methods & Clinical Development, 25, 158-169.

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Redox Reagents for Spectroscopic Analysis

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Titanium(III) chloride (20% w/v in 2 N HCl), titanium(IV) chloride (0.09 M in 20% HCl), sodium citrate (99%), potassium ferricyanide (99+%), 1,4-benzoquinone (99%), potassium ferrocyanide (98.5%), monobasic (98+%) and dibasic (≥99%) sodium phosphate, and sodium 2,6-dichlorophenol-indophenol (DCPIP, 98%) were acquired from ACROS Organics (Morris Plains, NJ). Sodium dithionite (>85%) was purchased from Alfa Aesar (Haverhill, MA). All chemicals were used as received.

Xin D., Xian M, & Chiu P.C. (2018). Chemical methods for determining the electron storage capacity of black carbon. MethodsX, 5, 1515-1520.

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Electrochemical Detection of Biomolecules

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Sodium nitrite
(Avocado Research Chemicals >95%), 4-aminobenzoic acid (Alfa Aesar
99%), hydrochloric acid (HCl) (Sigma-Aldrich 37%), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
hydrochloride (EDC) (Thermo Scientific 100%), N-hydroxysuccinimide
(NHS) (Thermo Scientific 100%), poly(propylene glycol)bis(2-aminopropyl
ether) (Sigma-Aldrich 100%), ethanolamine (Alfa Aesar 100%), β-cyclodextrin
(Acros Organics 98%), 3-mercaptopropionic acid (Acros Organics 99%),
ethanol (ACS Reagent), potassium ferrocyanide (Acros Organics, ACS
Reagent), potassium ferricyanide (Acros Organics, 98%), hydrocortisone
(98% Acros Organics), trans-resveratrol (Sigma-Aldrich
99%), acetaminophen (Sigma-Aldrich), uric acid (99% Alfa Aesar), and
artificial saliva (Pickering Laboratories In (pH 6.8)) were purchased.
PBS was prepared using a standard protocol from ThermoFisher Scientific
(pH 7.4). Raw, unprocessed pooled human urine from 20 donors was provided
by Lee BioSolutions (pH 6.11 and conductivity 13.06 mS/cm).
BASi gold and glassy carbon (GC) disk electrodes (area = 0.0201
cm²), gold-coated silicon wafers, and screen-printed carbon
electrodes were purchased from BASI, TED PELLA, and Metrohm DropSens,
respectively.
Ultra high purity (UHP) water is purified with
a reverse osmosis
and deionized with a Millipore Direct Q3 system and measured at 18.2
MΩ.

Panahi Z., Ren T, & Halpern J.M. (2022). Nanostructured Cyclodextrin-Mediated Surface for Capacitive Determination of Cortisol in Multiple Biofluids. ACS Applied Materials & Interfaces, 14(37), 42374-42387.

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Enzyme-Functionalized Electrochemical Biosensor

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All reagents were used as received without
further purification. Block copolymer poly(1,4-isoprene)-block-poly(ethylene oxide) PI-b-PEO (polydispersity:
1.01, Mn PI: 48, PEO: 12 kg mol^–1) was obtained
from Polymer Source. The following reagents were purchased from Merck:
glucose oxidase from Aspergillus niger (type X–S, lyophilized powder, 100.000–250.000 units
g^–1 solid without added oxygen), d-(+)-glucose
(≥99.5% (GC)), 1-butanol (99.4%), toluene (99.9%), toluene
(anhydrous, 99.8%), (3-glycidyloxypropyl)-trimethoxysilane (GLYMO)
(≥98%), aluminium tri-sec-butoxide (97%),
potassium chloride (KCl) (≥99.9%), (3-aminopropyl)triethoxysilane
(APTES) (99%), glutaraldehyde solution (Grade I, 25% in H₂O, specially purified for use as an electron microscopy fixative)
and ethanolamine hydrochloride (≥99.0%). Electrolytes potassium
ferricyanide (99+%, K₃[Fe(CN)₆]) and potassium
ferrocyanide (>98.5%, K₄[Fe(CN)₆]) were purchased
from ACROS Organics and Honeywell, respectively. Phosphate-buffered
saline (PBS) tablets were obtained from OXOID.

Jara Fornerod M.J., Alvarez-Fernandez A., Michalska M., Papakonstantinou I, & Guldin S. (2023). Glucose Oxidase Loading in Ordered Porous Aluminosilicates: Exploring the Potential of Surface Modification for Electrochemical Glucose Sensing. Chemistry of Materials, 35(18), 7577-7587.

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