MM 4-64 [N-(3-Triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide]
MM 4-64 (FM 4-64) is the abbreviation of our Membrane Marker 4-64, chemically called N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide. In literature this membrane marker is also called "FM 4-64" (FM® is the trademark of Molecular Probes). It is a lipophilic styryl dye that is used as a vital stain to follow bulk membrane-internalization and transport to the vacuole in yeast. MM 4-64 is a sensitive reporter of vacuolar dynamics, detecting such events as segregation structure formation during mitosis, vacuole fission/fusion events, and vacuolar morphology in different classes of vacuolar protein sorting mutants. It can be used for detecting endosome to vacuole membrane transport in vitro.
Example protocol
AT A GLANCE
Important
Expiration is 12 months from date of receiving.
SAMPLE EXPERIMENTAL PROTOCOL
- Make a 1-5 mM stock solution in DMSO.
- Prepare a working solution of 2-20 uM in ice-cold HHBS without Magnesium and calcium, and keep it on ice. Note: It is important to follow the temperature and time guidelines as closely as possible in order to slow down endocytosis and promote selective plasma membrane labeling and imaging. Endocytosis will likely occur within 10 minutes of staining. Note: HHBS (Hanks and Hepes Buffer) without magnesium and calcium is suggested for this protocol. The presence of magnesium or calcium speeds up endocytosis of the dye, resulting in poor plasma membrane selectivity.
- Remove the coverslip from the culture medium and quickly immerse it in the staining solution, on ice, for 1 minute. The plasma membranes will stain quickly.
- Remove the coverslip from the staining solution. Mount on a microscope slide, seal with paraffin, keep on ice, and image immediately in staining solution without washing.
Calculators
Common stock solution preparation
Table 1. Volume of Water needed to reconstitute specific mass of MM 4-64 [N-(3-Triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide] to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 164.606 µL | 823.032 µL | 1.646 mL | 8.23 mL | 16.461 mL |
5 mM | 32.921 µL | 164.606 µL | 329.213 µL | 1.646 mL | 3.292 mL |
10 mM | 16.461 µL | 82.303 µL | 164.606 µL | 823.032 µL | 1.646 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
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Citations
View all 22 citations: Citation Explorer
Quantitative Analysis of Rhodobacter sphaeroides Storage Organelles via Cryo-Electron Tomography and Light Microscopy
Authors: Parrell, Daniel and Olson, Joseph and Lemke, Rachelle A and Donohue, Timothy J and Wright, Elizabeth R
Journal: Biomolecules (2024): 1006
Authors: Parrell, Daniel and Olson, Joseph and Lemke, Rachelle A and Donohue, Timothy J and Wright, Elizabeth R
Journal: Biomolecules (2024): 1006
Essential role of Alix in regulating cardiomyocyte exosome biogenesis under physiological and stress conditions
Authors: Wang, Xinjian and Han, Shuxian and Liang, Jinxiu and Xu, Chen and Cao, Ranran and Liu, Shuoyang and Luan, Yi and Gu, Ying and Han, Peidong
Journal: Journal of Molecular and Cellular Cardiology (2024): 35--47
Authors: Wang, Xinjian and Han, Shuxian and Liang, Jinxiu and Xu, Chen and Cao, Ranran and Liu, Shuoyang and Luan, Yi and Gu, Ying and Han, Peidong
Journal: Journal of Molecular and Cellular Cardiology (2024): 35--47
Cell-lysis sensing drives biofilm formation in Vibrio cholerae
Authors: Prentice, Jojo A and van de Weerd, Robert and Bridges, Andrew A
Journal: Nature Communications (2024): 2018
Authors: Prentice, Jojo A and van de Weerd, Robert and Bridges, Andrew A
Journal: Nature Communications (2024): 2018
Synapsin condensation controls synaptic vesicle sequestering and dynamics
Authors: Hoffmann, Christian and Rentsch, Jakob and Tsunoyama, Taka A and Chhabra, Akshita and Aguilar Perez, Gerard and Chowdhury, Rajdeep and Trnka, Franziska and Korobeinikov, Aleksandr A and Shaib, Ali H and Ganzella, Marcelo and others,
Journal: Nature Communications (2023): 6730
Authors: Hoffmann, Christian and Rentsch, Jakob and Tsunoyama, Taka A and Chhabra, Akshita and Aguilar Perez, Gerard and Chowdhury, Rajdeep and Trnka, Franziska and Korobeinikov, Aleksandr A and Shaib, Ali H and Ganzella, Marcelo and others,
Journal: Nature Communications (2023): 6730
Fused expression of Sm1-Chit42 proteins for synergistic mycoparasitic response of Trichoderma afroharzianum on Botrytis cinerea
Authors: Liu, Hongyi and Wang, Shaoqing and Lang, Bo and Li, Yaqian and Wang, Xinhua and Chen, Jie
Journal: Microbial Cell Factories (2023): 1--15
Authors: Liu, Hongyi and Wang, Shaoqing and Lang, Bo and Li, Yaqian and Wang, Xinhua and Chen, Jie
Journal: Microbial Cell Factories (2023): 1--15
References
View all 41 references: Citation Explorer
A class Vb chitin synthase in Colletotrichum graminicola is localized in the growing tips of multiple cell types, in nascent septa, and during septum conversion to an end wall after hyphal breakage
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Authors: Nuriya M, Jiang J, Nemet B, Eisenthal KB, Yuste R.
Journal: Proc Natl Acad Sci U S A (2006): 786
Constitutive sharing of recycling synaptic vesicles between presynaptic boutons
Authors: Darcy KJ, Staras K, Collinson LM, Goda Y.
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Authors: Darcy KJ, Staras K, Collinson LM, Goda Y.
Journal: Nat Neurosci (2006): 315
DOC2A and DOC2B are sensors for neuronal activity with unique calcium-dependent and kinetic properties
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N-cadherin transsynaptically regulates short-term plasticity at glutamatergic synapses in embryonic stem cell-derived neurons
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Authors: Jungling K, Eulenburg V, Moore R, Kemler R, Lessmann V, Gottmann K.
Journal: J Neurosci (2006): 6968
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