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  3. DiD perchlorate

DiD is a long-chain carbocyanine dye. Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins.

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

DiD perchlorate Chemical Structure

DiD perchlorate Chemical Structure

CAS No. : 127274-91-3

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1 mg USD 35 In-stock
5 mg USD 92 In-stock
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Customer Review

Based on 4 publication(s) in Google Scholar

Top Publications Citing Use of Products
  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

DiD is a long-chain carbocyanine dye. Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins[2].

In Vitro

General Protocol
1. Preparing Stain Solutions of Di
1) Prepare DMF, DMSO or ethanol stock solutions: The stock solutions should be prepared in dimethyl formamide (DMF), dimethylsulfoxide (DMSO, or ethanol DMSO at 1-5 mM. DMF is preferable to ethanol as a solvent for Di. The stock solution should be used promptly. Any unused solution need to be aliquoted and refrozen at least -20℃. Avoid repeated freeze/thaw cycle. The solution can be stored for 6 months.
2) Prepare working solutions: Dilute the stock solutions into a suitable buffer such as serum-free culture medium, HBSS or PBS to make 1 to 5 μM working solutions. We do not recommend storing the aqueous solution for more than one day.
Note: The final concentration of the working solution should be empirically determined for different cell types and/or experimental conditions.
2. Suspension cells
1) 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
2) Add 1 mL of Di working solution, and then incubate at room temperature for 5-30 minutes.
3) Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
4) Wash twice with PBS, 5 minutes each time.
5) Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
3. Adherent cells
1) Culture adherent cells on sterile coverslips.
2) Remove the coverslip from the medium and aspirate the excess medium.
3) Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 minutes.
4) Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

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

In Vivo

Two weeks after injection of stained cells, a single, bright, DiD perchlorate (DiD)-positive cells located between 15 to 40 μm from the endosteum is found. Results reveal a progressive appearance of cell clusters of decreased dye intensity, consistent with the partitioning of DiD perchlorate label on cell division[3].

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

Molecular Weight

959.90

Formula

C61H99ClN2O4

CAS No.
Appearance

Solid

Emission (Em)

665

Excitation (Ex)

633

SMILES

CCCCCCCCCCCCCCCCCCN1C2=C(C=CC=C2)C(C)(C)/C1=C\C=C\C=C\C3=[N+](C4=C(C=CC=C4)C3(C)C)CCCCCCCCCCCCCCCCCC.O=Cl(=O)([O-])=O

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light, stored under nitrogen

*The compound is unstable in solutions, freshly prepared is recommended.

Solvent & Solubility
In Vitro: 

DMSO : 25 mg/mL (26.04 mM; ultrasonic and warming and heat to 60°C; 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 1.0418 mL 5.2089 mL 10.4178 mL
5 mM 0.2084 mL 1.0418 mL 2.0836 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.

  • 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: ≥ 1.67 mg/mL (1.74 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.67 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (16.7 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: ≥ 1.67 mg/mL (1.74 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.67 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (16.7 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).

*The compound is unstable in solutions, freshly prepared is recommended.

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: 98.96%

Dyeing Example
References
Animal Administration
[1]

1 to 5×105 cells per mL HSPCs are stained with 5 μM DiD perchlorate (DiD) or 7.5 μM DiI in PBS without serum for 10 min at 37°C, washed once in PBS and immediately injected into the tail vein of recipient mice. Unless stated differently, each imaged CD45.2 mouse receive 8,000 to 15,000 labelled CD45.1 HSPCs together with 3 to 5×105 CD45.2 supportive whole bone-marrow mononuclear cells to ensure survival[1].

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

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. The compound is unstable in solutions, freshly prepared is recommended.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.0418 mL 5.2089 mL 10.4178 mL 26.0444 mL
5 mM 0.2084 mL 1.0418 mL 2.0836 mL 5.2089 mL
10 mM 0.1042 mL 0.5209 mL 1.0418 mL 2.6044 mL
15 mM 0.0695 mL 0.3473 mL 0.6945 mL 1.7363 mL
20 mM 0.0521 mL 0.2604 mL 0.5209 mL 1.3022 mL
25 mM 0.0417 mL 0.2084 mL 0.4167 mL 1.0418 mL
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DiD perchlorate Related Classifications

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.

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DiD perchlorate
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