Hoechst 33258 trihydrochloride (Synonyms: bisBenzimide H 33258 trihydrochloride; H 33258 trihydrochloride)

Name: Hoechst 33258 trihydrochloride
Brand: MedChemExpress
SKU: HY-15558A
Rating: 5.0 (18 reviews)

Cat. No.: HY-15558A Purity: 99.51%: Data Sheet
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Hoechst 33258 trihydrochloride 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.

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

Hoechst 33258 trihydrochloride Chemical Structure

CAS No. : 23491-45-4

Size	Stock	Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
Solid
25 mg	In-stock	Estimated Time of Arrival: December 31
50 mg	In-stock	Estimated Time of Arrival: December 31
100 mg	In-stock	Estimated Time of Arrival: December 31
200 mg	Get quote
500 mg	Get quote

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Based on 18 publication(s) in Google Scholar

Other Forms of Hoechst 33258 trihydrochloride:

Hoechst 33258 In-stock

17 Publications Citing Use of MCE Hoechst 33258 trihydrochloride

: Hoechst 33258 trihydrochloride purchased from MedChemExpress. Usage Cited in: Virulence. 2022 Dec;13(1):444-457. [Abstract]; The apoptotic rate was detected by Hoechst staining (400×), HeLa cells were treated with siRNA for 24 h and followed by persistent infection.

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Biological Activity
Protocol
Purity & Documentation
References
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Description

IC₅₀ & Target

IC50: 51.31±4.56 μM (HeLa cell), 32.43±3.27 μM (HL60 cell), 15.42±2.16 μM (U937 cell)^[1]

Cellular Effect

Cell Line	Type	Value	Description	References
DU-145	IC₅₀	> 10 μM Compound: Hoechst 33258	Cytotoxicity against human DU145 cells after 48 hrs by SRB assay Cytotoxicity against human DU145 cells after 48 hrs by SRB assay	[PMID: 28513176]
HeLa	IC₅₀	23.52 μM Compound: Hoechst 33258	Antiproliferative activity against human HeLa cells after 48 hrs by MTT assay Antiproliferative activity against human HeLa cells after 48 hrs by MTT assay	[PMID: 30679134]
KG-1	IC₅₀	42 μM Compound: Hoechst 33258	Antiproliferative activity against human KG-1 cells assessed as inhibition of cell growth measured after 48 hrs by MTT assay Antiproliferative activity against human KG-1 cells assessed as inhibition of cell growth measured after 48 hrs by MTT assay	[PMID: 35152694]
MCF7	IC₅₀	48.1 μM Compound: Hoechst 33258	Antiproliferative activity against human MCF7 cells after 48 hrs by MTT assay Antiproliferative activity against human MCF7 cells after 48 hrs by MTT assay	[PMID: 30679134]

In Vitro

Preparation of Hoechst working solution
1.1 Preparation of stock solution
Take 10 mg and dissolve in 5 mL DMSO
Note: It is recommended that the stock solution be stored at 4℃ or - Store at 20℃ in the dark and avoid repeated freezing and thawing.
1.2 Preparation of Hoechst working solution
The diluted stock solution was dissolved in serum-free cell culture medium or PBS to obtain Hoechst working solution with a final concentration of 10 μg/mL.
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 for 3-5 min at 4℃ and discard the supernatant. Wash 2 times with PBS, 5 minutes each. The cell density was 1×10⁶/mL.
b. Add 1 mL of working solution and incubate at room temperature for 3-10 minutes.
c. Centrifuge at 400 g for 3-4 minutes at 4℃ and discard the supernatant.
d. Wash 2 times with PBS, 5 min each.
e. Resuspend cells in serum-free cell culture medium or PBS. Fluorescence microscopy or flow cytometry observation.
2.2 Adherent cells
a. Adherent cells were cultured on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium. c. Add 100 μL of working solution, shake gently to completely cover the cells, and incubate at room temperature for 3-10 minutes.
d. Wash 2 times with medium, 5 min each. Fluorescence microscope or flow cytometry observation.

Store
4°C, 1 year. Avoid light

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 medicine, household or other purposes.
3. For your safety and health, please wear a lab coat and disposable gloves for operation.

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

Molecular Weight

533.88

Formula

C₂₅H₂₇Cl₃N₆O

CAS No.

23491-45-4

Appearance

Solid

Color

Yellow to green

Emission (Em)

451

Excitation (Ex)

356

SMILES

OC1=CC=C(C2=NC3=CC=C(C4=NC5=CC=C(N6CCN(C)CC6)C=C5N4)C=C3N2)C=C1.[H]Cl.[H]Cl.[H]Cl

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°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 : 100 mg/mL (187.31 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H₂O : 12.5 mg/mL (23.41 mM; ultrasonic and warming and heat to 60°C)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	1.8731 mL	9.3654 mL	18.7308 mL
5 mM	0.3746 mL	1.8731 mL	3.7462 mL
10 mM	0.1873 mL	0.9365 mL	1.8731 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.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Molarity Calculator
Dilution Calculator

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

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

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
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: PBS
Solubility: 9.09 mg/mL (17.03 mM); Clear solution; Need ultrasonic and warming and heat to 60°C

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Calculation results:

Working solution concentration: mg/mL

Purity & Documentation

Purity: 99.51%

Select Batch:

Data Sheet (279 KB) SDS (394 KB)

COA (248 KB) HNMR (193 KB) LCMS (94 KB)

Handling Instructions (2659 KB)

References

[1]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through inductionof apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [Content Brief]

[2]. Stolić I, et al. Synthesis, DNA/RNA affinity and antitumour activity of new aromatic diamidines linked by 3,4-ethylenedioxythiophene. Eur J Med Chem. 2011 Feb;46(2):743-55. [Content Brief]

Cell Assay
^[2]

Hoechst 33258 is prepared as stock solutions in highly pure water. Working solutions in a concentration range of 10^-3-10^-6 mol/dm³ are prepared prior to testing. Cytotoxic effects of Hoechst 33258 on tested cell lines are determined by the MTT assay. Cells are seeded in 96 micro well flat bottom plates at a concentration of 2×10⁴ cells/mL and left overnight in the CO₂ incubator allowing them to attach to the plate surface. Growing medium is replaced with compound supplemented or control medium and incubated for 72 h. Fresh medium with 5 mg/mL of MTT is added onto cells and incubated for 4 h at 37°C. Upon media removal, water insoluble MTT-formazan crystals formed inside the living cells are dissolved in DMSO and the absorbance at 570 nm proportional to the number of living cells is measured on an Elisa Microplate Reader. All experiments are performed at least three times in triplicates.The GI₅₀ value, defined as the compound concentration (μM) leading to cellular growth inhibition by 50%, is calculated and used as a parameter to compare cytotoxicity among the compounds^[2].

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

References

[1]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through inductionof apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [Content Brief]

[2]. Stolić I, et al. Synthesis, DNA/RNA affinity and antitumour activity of new aromatic diamidines linked by 3,4-ethylenedioxythiophene. Eur J Med Chem. 2011 Feb;46(2):743-55. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

H₂O / DMSO	1 mM	1.8731 mL	9.3654 mL	18.7308 mL	46.8270 mL
	5 mM	0.3746 mL	1.8731 mL	3.7462 mL	9.3654 mL
	10 mM	0.1873 mL	0.9365 mL	1.8731 mL	4.6827 mL
	15 mM	0.1249 mL	0.6244 mL	1.2487 mL	3.1218 mL
	20 mM	0.0937 mL	0.4683 mL	0.9365 mL	2.3414 mL
DMSO	25 mM	0.0749 mL	0.3746 mL	0.7492 mL	1.8731 mL
	30 mM	0.0624 mL	0.3122 mL	0.6244 mL	1.5609 mL
	40 mM	0.0468 mL	0.2341 mL	0.4683 mL	1.1707 mL
	50 mM	0.0375 mL	0.1873 mL	0.3746 mL	0.9365 mL
	60 mM	0.0312 mL	0.1561 mL	0.3122 mL	0.7805 mL
	80 mM	0.0234 mL	0.1171 mL	0.2341 mL	0.5853 mL
	100 mM	0.0187 mL	0.0937 mL	0.1873 mL	0.4683 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Hoechst 33258 trihydrochloride 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:

Hoechst 33258 trihydrochloride23491-45-4bisBenzimide H 33258 trihydrochloride H 33258 trihydrochlorideFluorescent DyeHoechstDNAcellular dyeblue fluorescenceInhibitorinhibitorinhibit

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Hoechst 33258 trihydrochloride (Synonyms: bisBenzimide H 33258 trihydrochloride; H 33258 trihydrochloride)

Customer Review

Other Forms of Hoechst 33258 trihydrochloride:

Hoechst 33258 trihydrochloride Related Antibodies

17 Publications Citing Use of MCE Hoechst 33258 trihydrochloride

Featured Recommendations

•Related Small Molecules:

•Related Proteins:

Biological Activity

Protocol

Purity & Documentation

References

Customer Review

Hoechst 33258 trihydrochloride Related Antibodies

Molarity Calculator

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Complete Stock Solution Preparation Table

Hoechst 33258 trihydrochloride Related Classifications

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