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MM 4-64 [N-(3-Triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide]

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Physical properties
Molecular weight607.51
SolventWater
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


Molecular weight
607.51
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.

Platform


Fluorescence microscope

Excitation558 nm
Emission734 nm
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Important

Expiration is 12 months from date of receiving.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Make a 1-5 mM stock solution in DMSO.

  2. 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. 

  3. 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.

  4. 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 mg0.5 mg1 mg5 mg10 mg
1 mM164.606 µL823.032 µL1.646 mL8.23 mL16.461 mL
5 mM32.921 µL164.606 µL329.213 µL1.646 mL3.292 mL
10 mM16.461 µL82.303 µL164.606 µL823.032 µL1.646 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

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Citations


View all 20 citations: Citation Explorer
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
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
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
The Ypk1 protein kinase signaling pathway is rewired and not essential for viability in Candida albicans
Authors: Ram{\'\i}rez-Zavala, Bernardo and Kr{\"u}ger, Ines and Wollner, Andreas and Schwanfelder, Sonja and Morschh{\"a}user, Joachim
Journal: PLoS genetics (2023): e1010890
Resolving colistin resistance and heteroresistance in Enterobacter species
Authors: Doijad, Swapnil and Gisch, Nicolas and Frantz, Renate and Kumbhar, Bajarang and Falgenhauer, Jane and Imirzalioglu, Can and Falgenhauer, Linda and Mischnik, Alexander and Rupp, Jan and Behnke, Michael and others,
Journal: (2022)
A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of Pseudomonas aeruginosa
Authors: Arranz San Mart{\'\i}n, Alba and Drees, Steffen Lorenz and Fetzner, Susanne
Journal: Biomolecules (2022): 1656
The Bicarbonate Transporter (MoAE4) Localized on Both Cytomembrane and Tonoplast Promotes Pathogenesis in Magnaporthe oryzae
Authors: Dang, Yuejia and Wei, Yi and Zhang, Penghui and Liu, Xinchun and Li, Xinrui and Wang, Shaowei and Liang, Hao and Zhang, Shi-Hong
Journal: Journal of Fungi (2021): 955
Coevolution of eukaryote-like vps4 and ESCRT-III subunits in the asgard archaea
Authors: Lu, Zhongyi and Fu, Ting and Li, Tianyi and Liu, Yang and Zhang, Siyu and Li, Jinquan and Dai, Junbiao and Koonin, Eugene V and Li, Guohui and Chu, Huiying and others,
Journal: MBio (2020): e00417--20
Destructive role of TMAO in T-tubule and excitation-contraction coupling in the adult cardiomyocytes
Authors: Jin, Bu and Ji, Fangfang and Zuo, Anjun and Liu, Huiting and Qi, Lin and He, Yun and Wang, Qingyao and Zhao, Peng
Journal: International Heart Journal (2020): 355--363

References


View all 41 references: Citation Explorer
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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.
Journal: Nat Neurosci (2006): 315
DOC2A and DOC2B are sensors for neuronal activity with unique calcium-dependent and kinetic properties
Authors: Groffen AJ, Friedrich R, Brian EC, Ashery U, Verhage M.
Journal: J Neurochem (2006): 818
N-cadherin transsynaptically regulates short-term plasticity at glutamatergic synapses in embryonic stem cell-derived neurons
Authors: Jungling K, Eulenburg V, Moore R, Kemler R, Lessmann V, Gottmann K.
Journal: J Neurosci (2006): 6968
Yeast lipid rafts?--an emerging view
Authors: Wachtler V, Balasubramanian MK.
Journal: Trends Cell Biol (2006): 1
Sjl2p is specifically involved in early steps of endocytosis intimately linked to actin dynamics via the Ark1p/Prk1p kinases
Authors: Bottcher C, Wicky S, Schwarz H, Singer-Kruger B.
Journal: FEBS Lett (2006): 633
Actin-based motility during endocytosis in budding yeast
Authors: Kim K, Galletta BJ, Schmidt KO, Chang FS, Blumer KJ, Cooper JA.
Journal: Mol Biol Cell (2006): 1354
Activity-induced rapid synaptic maturation mediated by presynaptic cdc42 signaling
Authors: Shen W, Wu B, Zhang Z, Dou Y, Rao ZR, Chen YR, Duan S.
Journal: Neuron (2006): 401
A new photostable terrylene diimide dye for applications in single molecule studies and membrane labeling
Authors: Jung C, Muller BK, Lamb DC, Nolde F, Mullen K, Brauchle C.
Journal: J Am Chem Soc (2006): 5283
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
Authors: Amnuaykanjanasin A, Epstein L.
Journal: Protoplasma (2006): 155