Description

Hoechst 33258 is a cell-permeable, blue-fluorescent bis-benzimide dye widely used as a nucleic acid stain. Its high specificity for adenine-thymine (A-T) rich regions in the minor groove of double-stranded DNA makes it an essential tool for quantitative and qualitative analysis in molecular biology. This compound is critical for researchers and quality control laboratories in the biotechnology, pharmaceutical, and life sciences industries, where precise DNA quantification and visualization are required.

Application

• Fluorescent DNA Staining: For visualization of DNA in gel electrophoresis, particularly in agarose gels.
• Cell Biology & Cytometry: Staining of nuclear DNA in fixed cells for fluorescence microscopy and flow cytometry analysis.
• Quantitative DNA Assays: Used in fluorometric assays for determining DNA concentration in solution, often as an alternative to ethidium bromide.
• Chromosome Banding: Employed in cytogenetics for producing specific banding patterns on chromosomes.
• Apoptosis Detection: As a stain to identify apoptotic cells by characteristic nuclear condensation and fragmentation.
• Microbiological Research: For staining and enumerating bacteria in environmental or clinical samples.
• Quality Control in Bioprocessing: Monitoring DNA content and contamination in downstream purification processes.

Basic Information

Product Name	Hoechst 33258
CAS No.	23491-45-4
Molecular Formula	C25H24N6O · 3HCl
Molecular Weight	533.88 g/mol (as free base)
Synonyms	Bisbenzimide H 33258; HOE 33258; 2'-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi-1H-benzimidazole trihydrochloride; Pibenzimol; NSC 322921; Benzimide, bis, H 33258; DNA Stain Hoechst 33258
EINECS	Contact for details

Quality Control

Our Hoechst 33258 is produced and tested under strict quality management protocols to ensure batch-to-batch consistency and high performance in sensitive applications. Each lot undergoes comprehensive analysis, including identity confirmation, purity assessment, and functional performance testing in standard staining protocols. A detailed Certificate of Analysis (COA) documenting purity (typically ≥98% by HPLC), absorbance/fluorescence profiles, and suitability for biological staining is provided with every shipment.

Storage

Preserve in a tightly closed container, protected from light. Store at 2-8°C in a dry environment. The product is hygroscopic (moisture-sensitive) and light-sensitive; prolonged exposure to ambient conditions or light will degrade performance. For long-term stability, consider storage desiccated at -20°C.

Specification

Item	Specification
Appearance	Yellow to orange powder
Identification (IR)	Conforms to structure
Purity (HPLC)	≥ 98.0%
UV-Vis Absorption	λmax ~343 nm in water
Fluorescence Emission	λmax ~478 nm when bound to dsDNA
Water Content (KF)	≤ 5.0%
Solubility	Soluble in water, DMSO, DMF
Functional Test	Suitable for DNA staining

Note: Specifications can be tailored. Please contact us for the detailed technical data sheet of a specific grade.