Horse spleen apoferritin

Manufactured by Merck Group

Sourced in United States

Horse spleen apoferritin is a protein complex that can be used as a model system in various research applications. Apoferritin is the protein shell of the ferritin molecule, which functions as an iron storage and delivery protein. This product provides a source of the apoferritin protein without the iron core.

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Apoferritin (58 citations) Apoferritin from horse spleen (2 citations)

5 protocols using horse spleen apoferritin

Antioxidant Effects on Cell Cultures

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Cultures were used for experiments when greater than 11 days in vitro. Exclusion criteria were otherwise limited to bacterial or fungal contamination, cell peeling or cell death when examined under a phase contrast microscope prior to beginning the experiment. Unbiased treatment group assignments were guided by an online random number generator (www.random.org). DFO (Sigma-Aldrich, St. Louis, MO, USA) was added to cultures diluted in DMEM at 24 h before Hb or hydrogen peroxide treatment, reducing the serum content of the medium to 3.3% in the process. Control cultures were pretreated with DMEM alone. After DFO washout, exposure to human Hb A (Hemosol Inc, Etobicoke, Ontario, Canada) or hydrogen peroxide (CVS Health, Woonsocket, RI, USA) alone or with ascorbate (Spectrum Chemical Mfg Corp, New Brunswick, NJ, USA), Trolox (Sigma-Aldrich), DFO, or horse spleen apoferritin (Sigma-Aldrich) followed previously-published protocols [27 (link),28 (link)] in Minimal Essential Medium (Gibco 11430) containing 10 mM glucose (MEM10).
In one experiment, DFO was removed in stepwise fashion from cultures prior to Hb treatment. This was accomplished by reducing its concentration by half every 17 min for five cycles, followed by medium exchange. This 17 min interval approximates the plasma half-life of iron-free deferoxamine [29 ,30 ].

Peng D., Chen C.A., Ruhela D., Li Y, & Regan R.F. (2020). Deferoxamine deconditioning increases neuronal vulnerability to hemoglobin. Experimental cell research, 390(1), 111926.

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Apoferritin-Based Drug Delivery Protocol

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Horse spleen apoferritin, penicillin, streptomycin and triethylamine were purchased from Sigma Aldrich, (St. Louise, MO, USA); RPMI medium, phosphate buffered saline (PBS), and fetal bovine serum (FBS) and PrestoBlueTM reagent were from Thermo Fischer Scientific (Waltham, MA, USA). Van was from LC Laboratories (St. Woburn, MA, USA), acetonitrile from J. T. Baker (Phillipsburg, NJ, USA), and Iscove’s modified Dulbecco’s medium (IMDM) from Life Technologies (Carlsbad, CA, USA). All other chemicals were purchased from Lachner (Prague, Czech Republic). The purity of all chemicals met the standards of the American Chemical Society, unless noted otherwise.

Jáklová K., Feglarová T., Rex S., Heger Z., Eckschlager T., Hraběta J., Hodek P., Kolárik M, & Indra R. (2021). Apoferritin/Vandetanib Association Is Long-Term Stable But Does Not Improve Pharmacological Properties of Vandetanib. International Journal of Molecular Sciences, 22(8), 4250.

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Synthesis of Bismuth Nanoparticles

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The Dox was obtained from Sangon Biotech. Horse spleen apoferritin was obtained from Sigma-Aldrich. Bismuth nitrate and mannitol were obtained from Aladdin Industrial Co. Thioacetamide was obtained from J&K Chemical Co. Hydrochloric acid, glycine, and sodium hydroxide was obtained from Sinopharm Chemical Reagent Co. All chemicals were used as received without further purification.

Zhang Q., Chen J., Shen J., Chen S., Liang K., Wang H, & Chen H. (2019). Inlaying Radiosensitizer onto the Polypeptide Shell of Drug-Loaded Ferritin for Imaging and Combinational Chemo-Radiotherapy. Theranostics, 9(10), 2779-2790.

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Superoxide Anion Assay Using Apoferritin

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Horse spleen apoferritin, quinine sulfate, hydrogen peroxide (H₂O₂, 30 wt%), and 3,3′,5,5′-tetramethylbenzidine (TMB) were purchased form Sigma Aldrich (Shanghai, China). Superoxide anion assay kit was purchased from DOJINDO (China).

Shen Y., Zhang R, & Wang Y. (2023). One-pot hydrothermal synthesis of metal-doped carbon dot nanozymes using protein cages as precursors. RSC Advances, 13(10), 6760-6767.

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Encapsulation of Vincristine in Apoferritin

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The loading of VCR into the cavity of GKRK-APO and APO was prepared as described by previous report (Kilic et al., 2011 ), with minor modifications. Apoferritin from equine spleen (horse spleen apoferritin) was obtained from Sigma-Aldrich. Briefly, 100 μL of APO (10 mg) was dissolved in 0.15 M NaCl. Then, VCR was added into the solution at a final concentration of 1 mg/mL and stirred for 30 min. Subsequently, the pH of the mixed solution was adjusted to 2.0 using 0.1 M HCl and stirred for 10 min. In order to reassemble the protein to its native nanosphere form, the pH of the solution was then slowly increased to 7.4 by adding 0.1 M NaOH solution under constant stirring and continuous monitoring of pH through pH meter. The excessive VCR and the drug molecules outside of the protein shell were removed by dialysis against 0.9% NaCl solution. Finally, the VCR-loaded GKRK-APO and VCR-loaded APO were filtration sterilized by 0.2 μm filter and subpackaged to aseptic vials.

Zhai M., Wang Y., Zhang L., Liang M., Fu S., Cui L., Yang M., Gong W., Li Z., Yu L., Xie X., Yang C., Yang Y, & Gao C. (2018). Glioma targeting peptide modified apoferritin nanocage. Drug Delivery, 25(1), 1013-1024.

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