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  3. Fluorescent Dye
  4. HOE 32021

HOE 32021 is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution.

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HOE 32021 Chemical Structure

HOE 32021 Chemical Structure

CAS No. : 23623-06-5

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HOE 32021 Related Antibodies

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Description

HOE 32021 is a marker dye in Hoechst series. Hoechst is A live nuclear marker dye. Hoechst binds to the grooves in the DNA double strand, which tends to be A/ T-rich DNA strand. Although it binds to all nucleic acids, the A/ T-rich double strand DNA significantly enhances fluorescence intensity Therefore,Hoechst dye can be used for living cell labeling. The fluorescence intensity of Hoechst dye increases with the increase of pH of solution[1].

In Vitro

General Protocol
Preparation of Hoechst working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of in 5 mL DMSO
Note: It is recommended to store the stock solution at 4℃ or -20℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Hoechst working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain final concentration 10 μg/mL Hoechst working solution.
Note: Please adjust the concentration of Hoechst working solution according to the actual situation.
1.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 3-10 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 3-10 minutes.
d. Wash twice with medium, 5 minutes each time.Observation by fluorescence microscopy or flow cytometry.

Precautions
1. Please adjust the concentration of Hoechst working solution according to the actual situation.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

HOE 32021 Related Antibodies
Molecular Weight

438.52

Formula

C26H26N6O
CAS No.
Appearance

Solid

Color

Off-white to pink

Emission (Em)

451
Excitation (Ex)

356
SMILES

CN1CCN(C2=CC=C3N=C(C4=CC=C5N=C(C6=CC=C(OC)C=C6)NC5=C4)NC3=C2)CC1
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (114.02 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 2.2804 mL 11.4020 mL 22.8040 mL
5 mM 0.4561 mL 2.2804 mL 4.5608 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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

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This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo Dissolution Calculator
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation
References

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 (protect from 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 2.2804 mL 11.4020 mL 22.8040 mL 57.0099 mL
5 mM 0.4561 mL 2.2804 mL 4.5608 mL 11.4020 mL
10 mM 0.2280 mL 1.1402 mL 2.2804 mL 5.7010 mL
15 mM 0.1520 mL 0.7601 mL 1.5203 mL 3.8007 mL
20 mM 0.1140 mL 0.5701 mL 1.1402 mL 2.8505 mL
25 mM 0.0912 mL 0.4561 mL 0.9122 mL 2.2804 mL
30 mM 0.0760 mL 0.3801 mL 0.7601 mL 1.9003 mL
40 mM 0.0570 mL 0.2850 mL 0.5701 mL 1.4252 mL
50 mM 0.0456 mL 0.2280 mL 0.4561 mL 1.1402 mL
60 mM 0.0380 mL 0.1900 mL 0.3801 mL 0.9502 mL
80 mM 0.0285 mL 0.1425 mL 0.2850 mL 0.7126 mL
100 mM 0.0228 mL 0.1140 mL 0.2280 mL 0.5701 mL
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HOE 32021 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.

Keywords:

HOE 3202123623-06-5HOE32021HOE-32021Fluorescent DyeHoechstDNAcellular dyeblue fluorescenceInhibitorinhibitorinhibit

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