1. MCE Kits
  2. JC-1 Mitochondrial Membrane Potential Assay Kit

JC-1 Mitochondrial Membrane Potential Assay Kit 

Cat. No.: HY-K0601
Manual SDS

线粒体膜电位检测试剂盒 (JC-1) 是一种以 JC-1 为荧光探针,快速灵敏地检测组织、细胞或纯化的线粒体跨膜电势差的试剂盒,可以用于早期的细胞凋亡检测。

JC-1 Mitochondrial Membrane Potential Assay Kit

1.  客户无需承担相应的运输费用。

2.  同一机构(单位)同一产品试用装仅限申领一次,同一机构(单位)一年内

     可免费申领三个不同产品的试用装。

3.  试用装只面向终端客户

Size Price Stock Quantity
100 T ¥1660 In-stock

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  • Description

  • Storage

  • Protocol

  • Documentation

Description
& Advantages

ΔΨm, mitochondrial membrane potential, is an important parameter of mitochondrial function and has been used as an indicator of cell health. Variation of ΔΨm would be studied using JC-1. In healthy cells with high ΔΨm, JC-1 forms complexes known as J-aggregates. While in cells with low ΔΨm, JC-1 remains in the monomeric form. When excited at 490 nm, JC-1 monomers emit a green fluorescence with a maximum at ~520 nm. Aggregates of JC-1 emit an orange-red fluorescence with a maximum at ~590 nm.

Storage

Stored at -20°C protecting from light, and is stable for up to 12 months. Avoid repetitive freeze-thaw cycles while using. For immediate use, components may be stored at 2-8°C.

Protocol

1.   Labeling of Cells

a.   Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5× 105 cells/mL. Incubate the cells according to your normal protocol.

b.   Ensure that the JC-1 and DMSO has equilibrated to room temperature, and then prepare a 200 μM stock solution by dissolving the contents of one vial in 230 μL of the DMSO provided.

c.   For the control tube, allow the vial of CCCP has come to room temperature, add 1 μL of CCCP (50 mM). Incubate cells at 37°C for 5 minutes.

d.   Add 10 μL JC-1 (200 μM) per well to make the final concentration at 2 μM. Incubate cells at 37°C, 5% CO2, for 15-20 minutes. If additional labeling followed, for example with an annexin V, begin with step 2.a. If not, proceed with step 1.e.

e.   After incubation, centrifuge cells for 3-4 minutes at 400× g at 4°C, carefully aspirate the supernant.

f.   Wash cells twice with PBS (1×): add 2 mL PBS (1×) to suspend cells and vortex to mix thoroughly. Centrifuge cells for 3-4 minutes at 400× g at 4°C, carefully aspirate the supernant.

g.   Add 500 μL PBS (1×) to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader.

2.   Additional Labeling with Annexin V

a.   After step 1.d, wash cells twice with PBS (1×): add 2 mL PBS (1×) to suspend cells and vortex to mix thoroughly. Centrifuge cells for 3-4 minutes at 400× g at 4°C, carefully aspirate the supernant.

b.   Add 100 μL Annexin V binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4) to suspend the JC-1-stained cells.

c.   Add 5 μL Annexin V and incubate cells at 37°C, 5% CO2, for 15 minutes.

Note: 5 μL is appropriate for Annexin V, usage dose may vary according to different suppliers.

d.   Add 400 μL Annexin V binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4) to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader.

Documentation

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产品名称:
JC-1 Mitochondrial Membrane Potential Assay Kit
目录号:
HY-K0601
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