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> FM4-64 [162112-35-8]

FM4-64 [162112-35-8]

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Cat# HY-103466-1mg

Size : 1mg

Brand : MedChemExpress

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FM4-64 is a very lipophilic, water-soluble styrene dyes, can specifically bind to cell membranes and inner membrane organelles to produce fluorescence. FM4-64 is widely used in endocytic and exospic membrane structure markers.

For research use only. We do not sell to patients.

FM4-64 Chemical Structure

FM4-64 Chemical Structure

CAS No. : 162112-35-8

This product is a controlled substance and not for sale in your territory.

Based on 10 publication(s) in Google Scholar

Description

FM4-64 is a very lipophilic, water-soluble styrene dyes, can specifically bind to cell membranes and inner membrane organelles to produce fluorescence. FM4-64 is widely used in endocytic and exospic membrane structure markers.

In Vitro

1. Preparation of FM working solution
1.1 Preparation of the stock solution
Dissolve FM in DMSO to obtain 5 mM of FM.
Note: It is recommended to store the stock solution at -20℃or -80℃away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of FM working solution
Dilute the stock solution in HBSS to obtain 5-20 μM of FM working solution.
Note: Please adjust the concentration of FM working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells (6-well plate)
a. Centrifuge at 1000 g at 4℃for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL
b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4℃for 3-4 minutes and then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells, and then incubate at room temperature for 5-30 minutes.
d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.
Storage
-20℃,1 year.
Protect from light.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

FM4-64 Related Antibodies
Molecular Weight

607.51

Formula

C30H45Br2N3
CAS No.

162112-35-8
Appearance

Solid

Color

Gray to dark gray

Emission (Em)

634
Excitation (Ex)

558
SMILES

CC[N+](CCC[N+]1=CC=C(/C=C/C=C/C=C/C2=CC=C(N(CC)CC)C=C2)C=C1)(CC)CC.[Br-].[Br-]
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (82.30 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.6461 mL 8.2303 mL 16.4606 mL
5 mM 0.3292 mL 1.6461 mL 3.2921 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 1 mg/mL (1.65 mM); Clear solution

    This protocol yields a clear solution of ≥ 1 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 1 mg/mL (1.65 mM); Clear solution

    This protocol yields a clear solution of ≥ 1 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.94%

SDS (252 KB)

COA (243 KB) HNMR (281 KB) LCMS (149 KB)

Handling Instructions (2659 KB)
References

  • [1]. J E Gale, et al. FM1-43 dye behaves as a permeant blocker of the hair-cell mechanotransducer channel. J Neurosci. 2001 Sep 15;21(18):7013-25.  [Content Brief]

    [2]. Zal T, et al. Spectral shift of fluorescent dye FM4-64 reveals distinct microenvironment of nuclear envelope in living cells. Traffic. 2006;7(12):1607-1613.  [Content Brief]

    [3]. Stenovec M, et al. Distinct labelling of fusion events in rat lactotrophs by FM 1-43 and FM 4-64 is associated with conformational differences. Acta Physiol (Oxf). 2007;191(1):35-42.  [Content Brief]

    [4]. Bolte S, et al, Satiat-Jeunemaitre B. FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells. J Microsc. 2004;214(Pt 2):159-173.  [Content Brief]

  • [1]. J E Gale, et al. FM1-43 dye behaves as a permeant blocker of the hair-cell mechanotransducer channel. J Neurosci. 2001 Sep 15;21(18):7013-25.

    [2]. Zal T, et al. Spectral shift of fluorescent dye FM4-64 reveals distinct microenvironment of nuclear envelope in living cells. Traffic. 2006;7(12):1607-1613.

    [3]. Stenovec M, et al. Distinct labelling of fusion events in rat lactotrophs by FM 1-43 and FM 4-64 is associated with conformational differences. Acta Physiol (Oxf). 2007;191(1):35-42.

    [4]. Bolte S, et al, Satiat-Jeunemaitre B. FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells. J Microsc. 2004;214(Pt 2):159-173.

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.6461 mL 8.2303 mL 16.4606 mL 41.1516 mL
5 mM 0.3292 mL 1.6461 mL 3.2921 mL 8.2303 mL
10 mM 0.1646 mL 0.8230 mL 1.6461 mL 4.1152 mL
15 mM 0.1097 mL 0.5487 mL 1.0974 mL 2.7434 mL
20 mM 0.0823 mL 0.4115 mL 0.8230 mL 2.0576 mL
25 mM 0.0658 mL 0.3292 mL 0.6584 mL 1.6461 mL
30 mM 0.0549 mL 0.2743 mL 0.5487 mL 1.3717 mL
40 mM 0.0412 mL 0.2058 mL 0.4115 mL 1.0288 mL
50 mM 0.0329 mL 0.1646 mL 0.3292 mL 0.8230 mL
60 mM 0.0274 mL 0.1372 mL 0.2743 mL 0.6859 mL
80 mM 0.0206 mL 0.1029 mL 0.2058 mL 0.5144 mL
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FM4-64 Related Classifications

  • Others
  • Others
  • Fluorescent Dye
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

FM4-64162112-35-8Fluorescent Dyemembraneselectiveredfluorescentdyeendocytosisexocytosisorganelleorganization ,vesicleInhibitorinhibitorinhibit

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HY-103466-5mg
FM4-64 [162112-35-8]
 5mg