minute tm mitochondria isolation kit for mammalian cells and tissues  (Invent Biotechnologies)

Journal: Nature Communications

Article Title: CLPX acquires an iron-sulfur cluster to sustain mitochondrial proteostasis in cancer cells

doi: 10.1038/s41467-026-74080-2

Figure Lengend Snippet: Western blotting (WB) analyses of iron metabolism- and mitophagy-associated proteins in ( A , B ) Hepa1-6 allografts and ( C , D ) KYSE-150 xenografts revealed that iron deficiency treatment markedly decreased iron abundance and enhanced mitophagy level. E Flow cytometric analyses of Mtphagy Dye fluorescence intensity in the three cancer cell lines. F Transmission electron microscopic analyses of the three cancer cell lines exhibited engulfed mitochondria after iron deficiency treatment. Scale bar=1 μm. ( G , H) WB analyses of mitophagy-associated proteins in the three cancer cell lines before and after DFX treatment. WB analyses of TOMM20 and COX4I1 protein degradation in ( I ) HepG2, ( J ) Hepa1-6, and K KYSE-150 cells. ( L–N) Band intensity analysis results for ( I–K ). Fluorescence microscopic mitophagy assays utilizing the mito-Keima reporter plasmid for ( O ) HepG2, ( P ) Hepa1-6, and ( Q ) KYSE-150 cells. Mtphagy Dye, mitophagy detection dye; mito-Keima, mitochondria-targeted Keima fluorescent probe. These data collectively revealed that DFX treatment markedly enhanced mitophagy level in Hepa1-6 allografts, KYSE-150 xenografts, and the three cancer cell lines. Scale bar=10 μm. Data are presented as mean values ± standard deviation (SD), where the mean is represented by the measure of center and SD is represented the error bar (for B , D , E , L–N , which are the quantitative panels with error bars). For quantitative experiments ( E ), n = 5 independent biological replicates per experimental group; each biological replicate corresponds to an independent batch of cultured cancer cells (HepG2, Hepa1-6, or KYSE-150) or an independent animal xenograft/allograft (for A–D ) randomly assigned to control or DFX-treated conditions, and no technical replicates from the same cell batch or animal were pooled for statistical analysis. No technical replicates were used for statistical inference. Control groups consisted of PBS-treated or vehicle-treated cells/animals, and DFX-treated groups received deferasirox intervention. Representative WB ( A–D , G–N ), TEM ( F ), flow cytometry (panel E ), and fluorescence microscopy ( O–Q ) images are shown from three independent replicate experiments with consistent results; uncropped blots are provided in the Source Data. For all experiments, *, **, and *** respectively represent P values less than 0.05, 0.01 and 0.001 relative to the corresponding PBS or Control groups (unpaired two-sided Student’s t tests; exact P values are provided in the Source Data).

Article Snippet: For isolation of high-purity tissue mitochondria, we utilized the Mitochondria Isolation Kit (HY-K1061, MCE) for Tissue.

Techniques: Western Blot, Fluorescence, Transmission Assay, Plasmid Preparation, Standard Deviation, Cell Culture, Control, Flow Cytometry, Microscopy

Journal: MethodsX

Article Title: Purification of murine cortex mitochondria using a modified magnetic bead isolation method ☆

doi: 10.1016/j.mex.2026.103871

Figure Lengend Snippet: Final purified mitochondrial fractions stained with BioTracker 633 Red Mitochondria Dye (middle) and TOM20 (left) show viable pure mitochondria (merged; right). Scale bar is 5 µm.

Article Snippet: This method is based on Mitochondria Extraction Kit – Tissue and Mitochondria Isolation Kit mouse tissue, and their respective protocols by Miltenyi Biotec, with adjusted centrifugation conditions and added washing steps on final purified fractions.

Techniques: Purification, Staining