无论您使用哪种染料,四甲基罗丹明甲酯(TMRM)还是MitoTracker Red FM,未处理的细胞都会发出荧光。只要细胞线粒体膜电位降低就会导致荧光信号降低。最重要的是变化的程度。JC-1染料不仅强度改变,还有激发和发射比例光谱的改变。设置未处理的对照和用线粒体膜电位去稳定剂(如CCCP或FCCP)处理的阳性对照是非常重要的。这些染料仅用于活细胞,在固定处理的细胞中无法保留相同程度的信号。
I am seeing high background outside of my neuronal cells when using membrane potential indicators. What can I do to reduce background?
If you use our FluoVolt Membrane Potential Kit (Cat. No. F10488), the kit provides a background suppressor to reduce this problem. For other indicators, consider the use of BackDrop Background Suppressor (Cat no. R37603, B10511, and B10512).
Spatially organised mitochondrial calcium uptake through a novel pathway in chick neurones.
Authors:Coatesworth W, Bolsover S
Journal:Cell Calcium
PubMed ID:16338004
'A brief depolarisation of chick sensory neurones evokes a calcium increase in mitochondria that peaks 1-2s after the depolarisation event and then decays over tens of seconds. Peripheral mitochondria take up more calcium than do central ones, even when the cytosolic calcium increase is spatially homogeneous. The calcium influx into ... More
Tissue transglutaminase is a multifunctional BH3-only protein.
Authors:Rodolfo C, Mormone E, Matarrese P, Ciccosanti F, Farrace MG, Garofano E, Piredda L, Fimia GM, Malorni W, Piacentini M
Journal:J Biol Chem
PubMed ID:15485857
'Tissue transglutaminase (TG2) protein accumulates to high levels in cells during early stages of apoptosis both in vivo and in vitro. The analysis of the TG2 primary sequence showed the presence of an eight amino acid domain, sharing 70% identity with the Bcl-2 family BH3 domain. Cell-permeable peptides, mimicking the ... More
Methods for assessing autophagy and autophagic cell death.
Authors:Tasdemir E, Galluzzi L, Maiuri MC, Criollo A, Vitale I, Hangen E, Modjtahedi N, Kroemer G,
Journal:Methods Mol Biol
PubMed ID:18425442
'Autophagic (or type 2) cell death is characterized by the massive accumulation of autophagic vacuoles (autophagosomes) in the cytoplasm of cells that lack signs of apoptosis (type 1 cell death). Here we detail and critically assess a series of methods to promote and inhibit autophagy via pharmacological and genetic manipulations. ... More
Trafficking of ganglioside GD3 to mitochondria by tumor necrosis factor-alpha.
Authors:García-Ruiz C, Colell A, Morales A, Calvo M, Enrich C, Fernández-Checa JC
Journal:J Biol Chem
PubMed ID:12118012
'The interaction of mitochondria with proapoptotic proteins activates apoptosis pathways. Previous findings have identified ganglioside GD3 (GD3) as an emerging apoptotic lipid intermediate that targets mitochondria in response to death signals. Using immunoelectron and laser scanning confocal microscopy, we characterize the trafficking of GD3 to mitochondria in response to tumor ... More
Unreliability of the cytochrome c-enhanced green fluorescent fusion protein as a marker of cytochrome c release in cells that overexpress Bcl-2.
Authors:Unkila M, McColl KS, Thomenius MJ, Heiskanen K, Distelhorst CW
Journal:J Biol Chem
PubMed ID:11489892
'A cytochrome c-enhanced green fluorescent protein chimera (cyt-c.EGFP) was used to monitor the release of cytochrome c from mitochondria in Bcl-2-negative and Bcl-2-positive MDA-MB-468 breast cancer cells. A comparison was made with the intracellular distribution of endogenous cytochrome c based on Western blotting of cell fractions and immunocytochemistry. The release ... More