Table 1.
Contents and storage information.
JC-1 and JC-9 Mitochondrial Potential Sensors
Introduction
JC-1 and JC-9 are cationic dyes (Figure 1) that exhibit potential-dependent accumulation in mitochondria, indicated by a fluorescence emission shift from green (~525 nm) to red (~590 nm). Conquently, mitochondrial depolarization is indicated by a decrea in the red/green fluorescence intensity ratio. The potential-nsitive color shift is due to concentration-dependent formation of red fluorescent J-aggregates.1–3 JC-1 can be ud as an indicator of mitochondrial potential in a variety of
cell types, including myocytes 3 and neurons,4 as well as in intact tissues 5 and isolated mitochondria.6 JC-1 is more specific for mitochondrial versus plasma membrane potential, and more consistent in its respon to depolarization, than other cationic dyes such as DiOC 6(3) and rhodamine 123.7
The ratio of green to red fluorescence is dependent only on the membrane potential and not on other factors such as mitochondrial size, shape, and density that may influence
乌龟的功效与作用
single-component fluorescence signals. U of fluorescence ratio detection therefore allows rearchers to make comparative measurements of membrane potential and determine the percentage of mitochondria within a population that respond to an applied stimulus. Subtle heterogeneity in cellular respons can be discerned in this way.1,6 For example, four distinct patterns of mitochondrial membrane potential change in respon to glutamate receptor activation in neurons have been identified using confocal ratio imaging of JC-1 fluorescence.4 The most widely implemented application of JC-1 is for detection of mitochondrial depolar-ization occurring in the early stages of apoptosis (Figure 2).7–10
DMF
3
Typical staining protocols abstracted from the rearch literature are summarized in Table 2. Following incubation in dye-containing medium, it is usual to wash the cells before starting experimental obrvations.
早搏的症状和治疗方法•Figure 1. Structure of JC-1 (5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethyl-benzimidazolylcarbocyanine iodide).
+Cl
Cl
N N CH 23
CH 2
CH 3Cl
Cl CH
N N
CH 2CH
3CH 2
CH
3
CH
CH
I
2次元美女−
Optical Filters
A number of different optical filter configurations can be ud for analysis of JC-1 or JC-9 by
fluorescence microscopy (Table 3). For confocal lar scanning microscopy, the monomer
and J-aggregate forms can be excited simultaneously by 488 nm argon-ion lar sources. The
J-aggregate form can be excited lectively using the 568 nm argon-krypton lar line.
Appearance
Polarized mitochondria are marked by punctate orange-red fluorescent staining. On depolar-
有气势的团队歌曲ization, the orange-red punctate staining is replaced by diffu green monomer fluorescence.
Some of the green fluorescence may remain associated with mitochondria, due to potential-曹冲
independent interactions of the JC-1 monomer with mitochondrial membranes.2,3
Flow Cytometry
小兔子作文Staining
Typical staining protocols abstracted from the re-arch literature are summarized in Table 2.
Dissociated cells for flow cytometric analysis are diluted to a density of about 1 × 106 cells/mL
for staining.
Detector Configuration
When excited simultaneously by 488 nm argon-ion lar sources, the JC-1 monomer and
J-aggregate can be detected parately in the conventional flow cytometer FL1 and FL2
channels respectively (Figure 2).
Table 2. JC-1 cell staining conditions.
查阅的近义词
6. Human colon adenocarcinoma, J Cell Biol 138, 449 (1997);
7. Human premonocytic cell line, Proc Natl Acad Sci USA 93, 6458 (1996), Biochem Biophys Res Comm 197, 40 (1993);
8. J Physiol 486, 1 (1995).
Table 3. Optical filters for fluorescence microscope imaging of JC-1.
References
1. Proc Natl Acad Sci U S A 88, 3671 (1991);
2. Biochemistry 30, 4480 (1991);
3. J Physiol 486, 1 (1995);
4. J Neurosci 16, 5688 (1996);
5. Methods 18, 104 (1999);
6. Exp Cell Res 222, 84 (1996);
7. FEBS Lett 411, 77 (1997);
8. J Neurosci 18, 932 (1998);
9. J Cell Biol 138, 449 (1997); 10. Exp Cell Res 245, 170 (1998).
Product List Current prices may be obtained from our website or from our Customer Service Department.
丁香午月
Cat # Product Name Unit Size D22421 3,3’-dimethyl-α-naphthoxacarbocyan ine iodide (JC-9; DiNOC1(3)) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 mg T3168 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide (JC-1; CBIC2(3)) ......................................... 5 mg Contact Information
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