3,4-Dihydroxyhydrocinnamic acid was purchased from HEOWNS (China, Tianjin). Spermine (SPM) was purchased from Aladdin (China, Shanghai). MTT, 6-hydroxydopamine hydrochloride (6-OHDA) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Calcein AM was purchased from Abcam (Cambridge, UK). Deferoxamine mesylate (>98%) was purchased from MedChemexpress (New Jersey, USA). LDH Cytotoxicity Assay Kit, Annexin V-FITC Apoptosis Detection Kit, Reactive Oxygen Species Assay Kit, Lipid Peroxidation MDA Assay Kit, Total Superoxide Dismutase Assay Kit with WST-8, JC-1staining Kit were purchased from Beyotime (Nanjing, China). Hoechst 33,342 was purchased from Invitrogen (Carlsbad, CA, USA). Rabbit antibodies to Bcl-2, Bax, cytochrome C, Transferrin Receptor, Ferritin were purchased from Abcam (Cambridge, UK). Tyrosine hydroxylase (TH) and α-synuclein were purchased from Cell Signaling Technology (Beverly, MA, USA). Caspase-3, Caspase-9, PARP were purchased from Wanleibio. (China, Beijing). Ferroportin1 was purchased from Novus Biologicals (SanDiego, California, USA). β-Actin, α-tublin, hephaestin, DMT1 and HRP-conjugated goat anti-rabbit (mouse) IgG were purchased from Proteintech. (Rosemont, USA). Other chemicals and regents available were purchased form local commercial sources. PC12 cell line was gifted by professor Lu Ke at China Medical University.
Ferroportin 1
Manufactured by Novus Biologicals
Sourced in United States
Ferroportin 1 is a protein that functions as an iron exporter, regulating iron homeostasis in cells.
Automatically generated - may contain errors
Lab products found in correlation
Smad1, by Cell Signaling Technology (2 mentions)
Smad4, by Santa Cruz Biotechnology (2 mentions)
Ferritin, by Abcam (1 mentions)
Transferrin receptor, by Abcam (1 mentions)
Cytochrome c, by Abcam (1 mentions)
α synuclein, by Cell Signaling Technology (1 mentions)
Deferoxamine mesylate, by MedChemExpress (1 mentions)
Hrp conjugated goat anti rabbit mouse igg, by Proteintech (1 mentions)
Calcein am, by Abcam (1 mentions)
Caspase 9, by Wanlei (1 mentions)
Tyrosine hydroxylase, by Cell Signaling Technology (1 mentions)
α tublin, by Proteintech (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
β actin, by Proteintech (1 mentions)
Reactive oxygen species assay kit, by Beyotime (1 mentions)
Ldh cytotoxicity assay kit, by Beyotime (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (1 mentions)
Total superoxide dismutase assay kit with wst 8, by Beyotime (1 mentions)
Jc 1 staining kit, by Beyotime (1 mentions)
Lipid peroxidation mda assay kit, by Beyotime (1 mentions)
3 protocols using ferroportin 1
1
Iron Chelation and Neuroprotection
2
Immunoblotting Assay for Protein Analysis
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50–100 μg protein from liver, spleen or cell lysates were separated by 10–12% SDS-PAGE, and then transferred to nitrocellulose membranes. Primary antibodies used for immunoblotting assays were against DYKDDDDK tag (FLAG, Cell Signaling, cat. 2368S), HA-tag (Cell Signaling, cat. 2367S), Myc-tag (Cell Signaling, cat. 2276), α-tubulin (Ab frontier, cat. LF-MA0117A), p-SMAD1/5/8 (Cell Signaling, cat. 9511S), SMAD1/5/8 (Santa Cruz, cat. sc-6031-R), p-AMPK (Cell Signaling, cat. 2535S), AMPK (Cell Signaling, cat. 2532S), SHP (Santa Cruz, cat. sc-30169), SMAD4 (Santa Cruz, cat. sc-7966), SMAD1 (Cell Signaling, cat. 9743) and Ferroportin 1 (Novus Biologicals, cat. NBP1-21502).
3
Westerns for Protein Detection
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Cells were lysed in RIPA lysis buffer. Nuclear extracts were prepared with Nuclear Extract Kits (Active Motif, Carlsbad, CA, United States). Total cell lysates or nuclear and cytosol protein extracts were separated by 10% SDS–PAGE and blotted onto nitrocellulose membranes (Bio-Rad Laboratories). The membranes were immunoblotted with antibodies against the following: SMAD4 (1:500) (Santa Cruz Biotechnology, Santa Cruz, CA, United States), MyD88 (1:1000) (Cell Signaling, Danvers, MA, United States), phosphorylated SMAD5 (1:1000) (Abcam, Cambridge, MA, United States), SMAD1 (1:1000) (Cell Signaling), His (1:1000) (Genscript, Piscataway, NJ, United States), Flag (1:5000) (Genscript), HA (1:5000) (Genscript), Ubiquitin FK2 (1:100), Ferroportin 1 (1:1000) (Novus Biologicals, Littleton, CO, United States), GST (1:1000) (Genscript), and β-actin (1:10000) (Abcam, Cambridge, MA, United States). For secondary antibodies, anti–rabbit IgG (1:5000) or anti–mouse IgG (1:5000) were used. Antigen-antibody complexes were visualized with the ECL Western Blotting Detection Reagent (Invitrogen).
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