human transferrin elisa kit  (Multi Sciences (Lianke) Biotech Co Ltd)

Journal: Communications Biology

Article Title: HSPA4 restrains transferrin in dopaminergic neurons to attenuate ferroptosis in a Parkinson’s disease model

doi: 10.1038/s42003-025-08854-7

Figure Lengend Snippet: A HSPA4 colocalizes with transferrin in cells. The expression of HSPA4 (green) and transferrin (TF; red) in SH-SY5Y cells was analyzed by immunofluorescence analysis, and DAPI (blue) was used to stain the nucleus. Scale bar = 5 μm. B Intracellular transferrin was increased by HSPA4. SH-SY5Y cells were transfected with the plasmid expressing Hspa4 . Twenty-four hours post-transfection, the cells were subjected to immunofluorescence analysis using an anti-transferrin antibody. DAPI was used to stain the nucleus. Scale bar = 50 μm. C Quantification of the relative fluorescence intensity of transferrin, as shown in ( B ). n = 5 biologically independent samples. D HSPA4 sequesters transferrin in SH-SY5Y cells. SH-SY5Y cells were transfected with a plasmid expressing Hspa4 . Twenty-four hours post-transfection, the cells and cell culture media were collected, and HSPA4 expression was analyzed by western blot. Actin (left panel) and Coomassie blue staining (right panel) were used as loading controls. E HSPA4 reduces extracellular transferrin. SH-SY5Y cells were transfected with a plasmid expressing Hspa4 . Twenty-four hours post-transfection, the cell culture media were collected, and transferrin was measured with an ELISA kit. n = 4 biologically independent samples. F HSPA4 decreases the level of intracellular Fe 2+ . SH-SY5Y cells were transfected with a plasmid expressing Hspa4 . Twenty-four hours post-transfection, the cells were harvested for analysis of Fe 2+ . n = 3 biologically independent samples. EV: empty vector; TF: transferrin. All data are presented as means ± SEM. Statistical significance was determined by unpaired Student’s t -test.

Article Snippet: Transferrin in the cell culture medium was measured with a commercial ELISA kit (EK1201; MultiScience).

Techniques: Expressing, Immunofluorescence, Staining, Transfection, Plasmid Preparation, Fluorescence, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay