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 a critical tool for quantitative and qualitative analysis. This compound is essential for researchers and quality control laboratories in the biotechnology, pharmaceutical, and molecular diagnostics sectors, enabling applications from cell viability assays to chromosome banding.

Application

• Nucleic Acid Staining & Quantification: Used in fluorometric assays for determining DNA concentration in solution, often as an alternative to ethidium bromide.
• Cell Viability & Cytotoxicity Assays: Employed to stain the DNA of dead cells with compromised membranes, allowing for live/dead cell differentiation.
• Chromosome & Nuclear Staining: A key stain in cytogenetics for chromosome banding (e.g., Hoechst-banding) and fluorescence microscopy of nuclei.
• Flow Cytometry & Cell Cycle Analysis: Utilized to stain cellular DNA content for analyzing cell cycle phases (G0/G1, S, G2/M) by flow cytometry.
• Fluorescence In Situ Hybridization (FISH): Acts as a counterstain to visualize chromosomes alongside specific FISH probes.
• Microbiological Research: Applied for staining bacterial DNA and for studying biofilms under fluorescence microscopy.

Basic Information

Product Name	Hoechst 33258
CAS No.	23491-44-3
Molecular Formula	C25H24N6O · 3HCl · 3H2O
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; Fluorescent DNA Stain; Nuclear Stain 33258; AT-selective DNA minor groove binder
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 biological assays. Each lot is accompanied by a comprehensive Certificate of Analysis (COA) detailing purity, identity, and functional performance data. We support compliance with relevant research and quality control standards.

Storage

Preserve in a tightly closed container, protected from light. Store in a cool, dry place at 2-8°C. This product is hygroscopic (moisture-sensitive) and strictly light-sensitive. Allow the vial to equilibrate to room temperature before opening to minimize moisture uptake.

Specification

Item	Specification
Appearance	Yellow to orange powder
Identification (IR)	Conforms to structure
Purity (HPLC)	≥ 98.0%
Water Content (KF)	≤ 5.0%
Fluorescence Qualification	Suitable for cell staining

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