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Apoferritin

Manufactured by Merck Group
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Sourced in United States, Germany, United Kingdom

Apoferritin is a protein-based nanomaterial used as a lab equipment component. It functions as a versatile nanocarrier, capable of encapsulating various molecules and materials within its hollow structure.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (16 mentions) β amylase, by Merck Group (13 mentions) Carbonic anhydrase, by Merck Group (13 mentions) Alcohol dehydrogenase, by Merck Group (13 mentions) Thyroglobulin, by Merck Group (12 mentions) Blue dextran, by Merck Group (9 mentions) 70s ribosomes, by New England Biolabs (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Deferoxamine, by Merck Group (3 mentions) Ferritin, by Merck Group (3 mentions) Superose 12 hr 10 30 column, by GE Healthcare (3 mentions) Vitamin b12, by Merck Group (3 mentions) Apo transferrin, by Merck Group (2 mentions) 40 m cell strainer, by BD (2 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions) Dmem medium, by Corning (2 mentions) Biliverdin, by Frontier Scientific (2 mentions) Murine recombinant il 4, by Miltenyi Biotec (2 mentions) Murine recombinant ifn γ, by Roche (2 mentions) M csf, by Miltenyi Biotec (2 mentions)

Close spelling variants of this product

Horse spleen apoferritin (5 citations) Apoferritin from equine spleen (3 citations) Apoferritin and ferritin (2 citations) Apoferritin from horse spleen (2 citations) Equine Apoferritin (2 citations)

58 protocols using apoferritin

1

Therapeutic Targeting of Inflammation

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Apoferritin: Purified horse spleen ferritin (Sigma Ref: F4503) or Apoferritin (Sigma Ref: A3641) in sterile 0.9% saline was administered (i.p.; 200 μL in saline) 24 hr before or 6 hr after CLP. Vehicle (0.9% saline) or bovine serum albumin (BSA, Sigma Ref: A3059) at the same dosage were administered to control mice. D-glucose (Sigma Ref: G-8270) (200 μL 2 mg/g BW in water) or Pyruvate (Sigma Ref: P4562) (200 μL 4 mg/g BW in water) were administered via gavage twice daily for 4 days starting 12 hr after CLP and then once daily for 3 days. N-acetylcystein (NAC) was administered (i.p.; 200 μL, 15mg/kg in sterile PBS) twice daily for 12 consecutive days starting 48 hr before CLP. Butylated Hydroxianisole (BHA; Sigma, ref: B1253) was administered (gavage.; 100 μL, 50mg/kg in corn oil) twice daily for 12 consecutive days starting 48 hr before CLP. Hemin (Frontier Scientific Ref: H651-9) or empty protoporphyrin (Frontier Scientific Ref: P562) were administered to C57BL/6, Tlr4−/− and Ifnr1−/− (i.p.; 30 mg/kg) or to Fthlox/lox and Mx1CreFthΔ/Δ (i.p.; 25-30 mg/kg). Poly(I:C) was administered to G6pclox/lox and to AlbCreERT2G6pc1Δ/Δ (intra-retro orbitally; 30 mg/kg BW).
Weis S., Carlos A.R., Moita M.R., Singh S., Blankenhaus B., Cardoso S., Larsen R., Rebelo S., Schäuble S., Del Barrio L., Mithieux G., Rajas F., Lindig S., Bauer M, & Soares M.P. (2017). Metabolic Adaptation Establishes Disease Tolerance to Sepsis. Cell, 169(7), 1263-1275.e14.
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2

Apoferritin and Ribosomes Sample Preparation

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Apoferritin was purchased from Sigma Aldrich, 400 kDa, A3660, 2.3 mg/ml. Protein solution stored in 50% glycerol was exchanged into a cryo compatible buffer (50 mM Tris-Cl [pH 7.6]; 150 mM NaCl) using Amicon Ultra-15 centrifugal filter units (100 kDa cutoff membrane). 70S ribosomes were purchased from New England BioLabs Inc, 2MDa. Protein solution was stored in 20 mM HEPES-KOH [pH 7.6], 10 mM Mg(CH3COO)2, 30 mM KCl, and 7 mM β-mercaptoethanol after diluting the sample to 1 mg/ml from 33.3 mg/ml.
Dandey V.P., Budell W.C., Wei H., Bobe D., Maruthi K., Kopylov M., Eng E.T., Kahn P.A., Hinshaw J.E., Kundu N., Nimigean C.M., Fan C., Sukomon N., Darst S.A., Saecker R.M., Chen J., Malone B., Potter C.S, & Carragher B. (2020). Time-resolved cryoEM using Spotiton. Nature methods, 17(9), 897-900.
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3

Preparation and Storage of Biological Samples

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Bovine vitreous was purchased from InVision Bioresources, Seattle, Washington, USA. It was frozen after harvesting and shipped on dry ice. For use in experiments, it was quickly thawed, homogenized while ice-cold, and sterile-filtered. Aliquots were then stored at −80°C until used.
Human Hb A was obtained as a gift from Hemosol, Inc, Etobicoke, Ontario, Canada.
Apotransferrin was purchased from Millipore-Sigma, Burlington, MA, USA and apoferritin was purchased from Sigma-Aldrich, St Louis, MO, USA.
Hyaluronic acid was purchased from R&D Systems, Minneapolis, MN, USA.
Culture media (MEM and DMEM) were purchased from Gibco-Thermo Fisher Scientific, Gaithersburg, MD, USA; serum was purchased from GE Healthcare Hyclone, Logan, UT, USA. Other experimental reagents were purchased from Sigma-Aldrich unless otherwise indicated.
Chen-Roetling J., Regan K.A, & Regan R.F. (2018). Protective Effect of Vitreous against Hemoglobin Neurotoxicity. Biochemical and biophysical research communications, 503(1), 152-156.
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4

NiV G-EphrinB2 Binding Kinetics

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EphrinB2 protein was mixed with 10 μg of NiV G ectodomain in 100 μl of 50 mM tris (pH 7.5) and 150 mM NaCl at the indicated molar ratio. Samples were injected at 100-μl injection volumes and run at 0.2 ml/min on a S200 5/150 GL column with 3-ml bed volume. Sizing standards were blue dextran (2000 kDa, void volume), thyroglobulin (664 kDa), apoferritin (443 kDa), β-amylase (200 kDa), and BSA (66 kDa) (Sigma-Aldrich).
Wong J.J., Chen Z., Chung J.K., Groves J.T, & Jardetzky T.S. (2021). EphrinB2 clustering by Nipah virus G is required to activate and trap F intermediates at supported lipid bilayer–cell interfaces. Science Advances, 7(5), eabe1235.
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5

Isolation and Polarization of Murine Bone Marrow-Derived Macrophages

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BMDMs were isolated and cultured as previously described with minor modifications (38 (link)). Mouse bone marrow cells were flushed from the femurs of mice and passed through a 40-µm cell strainer (BD Falcon), and red blood cells were lysed in ACK buffer (0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA). Remaining cells were incubated in DMEM medium (Corning Cellgro) containing 10% FBS (Atlanta Biologicals), 30 ng/ml MCSF (Miltenyi Biotec) and 1% penicillin/streptomycin (Life Technologies). BMDM were cultured for 7 days and the purity of the culture was determined to be >85% based on analysis of CD11b expression. Polarization of cells was performed using 100 U/ml murine recombinant IFN γ (Roche) or 20 ng/ml murine recombinant IL-4 (Miltenyi Biotec). For studies involving HO by-products or ferritin, pre-treatment was performed 16 hours prior to the addition of cytokines. Macrophages were pre-treated with biliverdin (Frontier Scientific), bilirubin (Frontier Scientific), deferoxamine (Sigma), CORM (Sigma) or Apoferritin (Sigma). Recombinant H ferritin and L ferritin were generously provided by Dr. Arosio. HO-1 overexpressing macrophages were isolated from humanized HO-1 BAC transgenic mice (81 (link)). Gene expression analysis was performed at 4 hours and protein expression analysis was performed at 24 hours after cytokine treatment.
Bolisetty S., Zarjou A., Hull T.D., Traylor A., Perianayagam A., Joseph R., Kamal A.I., Arosio P., Soares M.P., Jeney V., Balla J., George J.F, & Agarwal A. (2015). Macrophage and epithelial cell H-ferritin expression regulates renal inflammation. Kidney international, 88(1), 95-108.
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6

Size-exclusion Chromatography of rhGFAT2 Protein

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The purified rhGFAT2-his protein was subjected to a size-exclusion chromatography using a Superdex 200 column (GE Healthcare, USA). The column was equilibrated with 1 CV of 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM DTT, 0.5% NP-40 prior to sample loading at a flow rate of 1 ml/min. The fractions were collected and analyzed by SDS-PAGE. The molecular weight of rhGFAT2 oligomer was estimated according to the retention time of standard proteins (Thyroglobulin—669 kDa, Apoferritin—443 kDa, β-amilase—200 kDa, BSA—66 kDa, Carbonic anhydrase—29 kDa, and Citocrome C oxidase—12.4 kDa) acquired from Sigma Co.
Oliveira I.A., Allonso D., Fernandes T.V., Lucena D.M., Ventura G.T., Dias W.B., Mohana-Borges R.S., Pascutti P.G, & Todeschini A.R. (2020). Enzymatic and structural properties of human glutamine:fructose-6-phosphate amidotransferase 2 (hGFAT2). The Journal of Biological Chemistry, 296, 100180.
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7

Isolation and Polarization of Murine Bone Marrow-Derived Macrophages

Cited 1 time
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BMDMs were isolated and cultured as previously described with minor modifications (38 (link)). Mouse bone marrow cells were flushed from the femurs of mice and passed through a 40-µm cell strainer (BD Falcon), and red blood cells were lysed in ACK buffer (0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA). Remaining cells were incubated in DMEM medium (Corning Cellgro) containing 10% FBS (Atlanta Biologicals), 30 ng/ml MCSF (Miltenyi Biotec) and 1% penicillin/streptomycin (Life Technologies). BMDM were cultured for 7 days and the purity of the culture was determined to be >85% based on analysis of CD11b expression. Polarization of cells was performed using 100 U/ml murine recombinant IFN γ (Roche) or 20 ng/ml murine recombinant IL-4 (Miltenyi Biotec). For studies involving HO by-products or ferritin, pre-treatment was performed 16 hours prior to the addition of cytokines. Macrophages were pre-treated with biliverdin (Frontier Scientific), bilirubin (Frontier Scientific), deferoxamine (Sigma), CORM (Sigma) or Apoferritin (Sigma). Recombinant H ferritin and L ferritin were generously provided by Dr. Arosio. HO-1 overexpressing macrophages were isolated from humanized HO-1 BAC transgenic mice (81 (link)). Gene expression analysis was performed at 4 hours and protein expression analysis was performed at 24 hours after cytokine treatment.
Bolisetty S., Zarjou A., Hull T.D., Traylor A., Perianayagam A., Joseph R., Kamal A.I., Arosio P., Soares M.P., Jeney V., Balla J., George J.F, & Agarwal A. (2015). Macrophage and epithelial cell H-ferritin expression regulates renal inflammation. Kidney international, 88(1), 95-108.
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8

Modulating Iron Homeostasis in LECs

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Freshly isolated primary LECs were purchased from Cell Biologics (CD1017) and cultured on gelatin in complete growth media containing 5% fetal calf serum (Cell Biologics M1168) at 37 °C in a 5% CO2 95% air atmosphere. BMP6 induction experiments were performed up to passage 4–8 to maintain iron-mediated BMP6 responsiveness.
For iron depletion experiments, cells were treated with complete media supplemented with 100 μM deferoxamine (Sigma D9533) for 24 hours. For iron loading experiments, cells were serum starved for 8–16 hours prior to treatment with 200 μg/mL ferric ammonium citrate (Sigma F5879) for 6–24 hours, 30 μM holo-transferrin (Sigma T0665) or apo-transferrin (Sigma T1147) for 24 hours, or 100 nM holo-ferritin (Sigma F4503) or apo-ferritin (Sigma 178440) for 24 hours.
For gene knockdown experiments, cells were transfected with 40 nM non-targeting control siRNA (Invitrogen 4390846) or siRNA targeting Tfrc (Invitrogen s75457) using Lipofectamine RNAiMAX (Invitrogen) per the manufacturer’s instructions for 48 hours prior to iron treatment as described above. Gene knockdown was confirmed by qRT-PCR and immunoblotting.
Fisher A.L., Wang C.Y., Xu Y., Joachim K., Xiao X., Phillips S., Moschetta G.A., Alfaro-Magallanes V.M, & Babitt J.L. (2022). Functional role of endothelial transferrin receptor 1 in iron sensing and homeostasis. American journal of hematology, 97(12), 1548-1559.
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9

Gelatin Molecular Mass Analysis

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Using a fast protein liquid chromatography (GE Healthcare Europe GmbH, Munich, Germany) equipped using a silica gel packed in a TSKgel G2000SWXL column (7.8 mm id × 30 cm), the MM analysis of gelatin was done as previously reported [23 ]. Elution was performed using 0.1 M phosphate buffer; 0.2 M sodium chloride. The standard markers were blue dextran (2000 kDa), apoferritin (443 kDa), β-amylase (200 kDa) and bovine serum albumin (66 kDa) (Sigma-Aldrich). The gelatins and standard markers were loaded into the column at 5 mg/ml. The MM was calculated from the standard curve.
Salem A., Abdelhedi O., Sebii H., Ben Taheur F., Fakhfakh N., Jridi M., Zouari N, & Debeaufort F. (2023). Techno-functional characterization of gelatin extracted from the smooth-hound shark skins: Impact of pretreatments and drying methods. Heliyon, 9(9), e19620.
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10

Synthesis and Characterization of MBA-Conjugated Apoferritin

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Apoferritin and CuCl2·2H2O were purchased from Sigma-Aldrich (MO, USA). Na2S was purchased from Sinopharm Chemical Reagent Co., Ltd. MBA (MW = 768) was synthesized in our laboratory. 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were purchased from Bide Pharmatech Ltd. (Shanghai, China). 3-(4,5-Dimethylthialzol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), RPMI-1640, fetal bovine serum (FBS), penicillin, streptomycin, and trypsin–EDTA were purchased from Gibco (Life Technologies, Shanghai, China). The reagents in this study were used without further purification.
He Y., Shen Y., Zhou S., Wu Y., Yuan Z., Wei C., Gui L., Chen Y., Gu Y, & Chen H. (2018). Near infrared dye loaded copper sulfide-apoferritin for tumor imaging and photothermal therapy. RSC Advances, 8(26), 14268-14279.
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