Hoechst 33258 staining solution - BioReagent, Biological Stain, Biological dye grade, Suitable for Immunofluorescence(IF), for fluorescence analysis, for cell culture, for microscopy, ready-to-use, 1.0 mg/mL in H₂O , CAS No.23491-45-4

CAS: 23491-45-4 Cat. No.: H1373484 Molecular Weight: 533.90 Beilstein Registry Number: 4088183 EC Number: 245-690-6 PubChem CID: 31953

GRADE & PURITY BioReagent, Ready-to-use, Biological Stain, for Fluorescence analysis, Biological dye grade, for Microscopy, for Cell culture, Suitable for Immunofluorescence(IF), 1.0 mg/mL in H₂O

Synonyms

Hoechst 33258 | Hoechst 33258 Stain Solution | Hoechst 33258 Stain Solution(1mg/ml) | Hoechst 33258 Staining Dye Solution | Hoechst 33258 Solution (1 mg/mL)

Storage

Protected from light,Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Application

Cell Apoptosis Analysis, Cell Cycle Analysis, Cell Stain, Flow Cytometry, IF/ICC

Overview

Hoechst 33258 is a blue to blue-green fluorescent live cell dye that can label DNA. Hoechst 33258 can specifically bind to the DNA groove (tending to bind to DNA rich in A/T at the same time), resulting in a significant increase in fluorescence intensity. Hoechst 33258 can pass through the cell membrane, causing changes in DNA structure, such as G2/M phase arrest. Hoechst 33258 can bind to live cells or fixed cells, and the fluorescence intensity increases with the increase of solution pH. As a DNA-specific probe, Hoechst 33258 can be used for detecting DNA content and analyzing cell cycle, etc. The excitation wavelength of Hoechst 33258 is 350-365 nm, and the emission wavelength is 460-490 nm.

The concentration of this product is 1.0 mg/mL. When used for nuclear staining, the recommended working concentration is 1-10 μg/mL.

Instruction for use 

1.For fixed cells or tissues:

a. The cells were inoculated onto the culture plates/flasks and cultured until they reached the logarithmic growth phase. If cell fixation was required, the cells were fixed with 4% paraformaldehyde at room temperature for 10-15 minutes, then washed twice with PBS, each time for 5 minutes.

b. Add the Hoechst 33258 working solution and incubate at 37°C in the dark for 10-30 minutes.

c. Remove the Hoechst 33258 staining solution and wash the cells twice with PBS for 5 minutes each time.

2.For living cells or tissues：

a. Collect the cells, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant.

b. Resuspend the cells in PBS, add the Hoechst 33258 working solution, and incubate at room temperature in the dark for 15-20 minutes.

c. Centrifuge at 1000 rpm for 5 minutes, discard the staining solution, wash twice with PBS for 5 minutes each time; then resuspend the cells in PBS.

d.Observe directly under a fluorescence microscope or under a fluorescence microscope after mounting. When cells undergo apoptosis, the nuclei of apoptotic cells will show dense and compact staining, or fragmented and dense staining.

Matters needing attention

1.Depending on the application of Hoechst 33258 staining solution, it is recommended to perform a titration before use to determine the optimal working concentration.

2.Fluorescent dyes all have the problem of quenching. It is recommended to complete the detection as soon as possible after staining. Live cells or tissues should be observed immediately after staining.

3.This product is only for professional scientific research use. It shall not be used for clinical diagnosis or treatment, nor for food or medicine. It shall not be stored in ordinary residences.

4.For your safety and health, please wear laboratory coats and disposable gloves during operation.

Specifications

Synonyms

Hoechst 33258 | Hoechst 33258 Stain Solution | Hoechst 33258 Stain Solution(1mg/ml) | Hoechst 33258 Staining Dye Solution | Hoechst 33258 Solution (1 mg/mL)

Specifications & Purity

BioReagent, Biological Stain, Biological dye grade, Suitable for Immunofluorescence(IF), for fluorescence analysis, for cell culture, for microscopy, ready-to-use, 1.0 mg/mL in H₂O

Stability And Storage

Store at -20℃ long term (12 months). Store in the dark. Avoid repeated freezing and thawing.

Storage

Protected from light, Store at -20°C, Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

Biological dye grade, Biological Stain, BioReagent, for Cell culture, for Fluorescence analysis, for Microscopy, Ready-to-use, Suitable for Immunofluorescence(IF)

Images

Cell Staining: Hoechst 33258 Staining Solution (H1373484)

Hoechst 33258 Staining Solution (H1373484) - Cell Staining 

Apoptosis was induced in Jurkat cells by a 16-hour treatment with 2 μM camptothecin (C111281) and assessed using Hoechst 33258 staining solution (H1373484). This dye exhibits enhanced binding to condensed and fragmented chromatin in apoptotic cells, resulting in intense blue fluorescence under confocal laser scanning microscopy. In contrast, viable cells show only weak, diffuse nuclear fluorescence.

Names and Identifiers

• Isomeric SMILES: CN1CCN(CC1)C2=CC3=C(C=C2)N=C(N3)C4=CC5=C(C=C4)N=C(N5)C6=CC=C(C=C6)O.Cl.Cl.Cl
• WGK Germany: 3
• RTECS: SM1140500
• PubChem CID: 31953
• Molecular Weight: 533.90
• Beilstein: 4088183

Application

• Application: Dilution info
• Flow Cytometry:

  1-10 μg/ml ;

Chemical and Physical Properties

• Sensitivity: Light-sensitive
• Melt Point(°C): 314°C

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