Synonym: 2′-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5′-bi(1H-benzimidazole) trihydrochloride; 2-[2-(4-Hydroxyphenyl)-6-benzimidazoyl]-6-(1-methyl-4-piperazyl)benzimidazole trihydrochloride; BBIH; BXI-72; HOE 33258; Hoechst 33258; NSC334072
CAS Number: 23491-45-4
Empirical Formula (Hill Notation): C25H24N6O · 3HCl
Molecular Weight: 533.88
EC Number: 245-690-6
MDL Number: MFCD00012679
Linear Formula: C25H24N6O · 3HCl
Product Type: Chemical
| assay |
≥98.0% (HPLC) |
| fluorescence |
λex 338 nm; λem 505 nm (pH 7.0) |
| |
λex 355 nm; λem 465 nm in TE buffer; DNA |
| form |
powder |
| grade |
for fluorescence |
| InChI |
1S/C27H28N6O.3ClH.H2O/c1-32-10-12-33(13-11-32)22-5-9-25-21(15-22)17-29-27(31-25)19-4-8-24-20(14-19)16-28-26(30-24)18-2-6-23(34)7-3-18;;;;/h2-9,14-15,34H,10-13,16-17H2,1H3,(H,28,30)(H,29,31);3*1H;1H2 |
| InChI key |
OWRSPPSBNWJJAR-UHFFFAOYSA-N |
| Quality Level |
200  |
| SMILES string |
Cl[H].Cl[H].Cl[H].[H]O[H].CN1CCN(CC1)c2ccc3NC(=NCc3c2)c4ccc5NC(=NCc5c4)c6ccc(O)cc6 |
| solubility |
ethanol: soluble |
| |
H2O: soluble |
| |
water and ethanol: 10 mg/mL |
| storage temp. |
2-8°C |
| Application: |
A bibenzimidazole dye that has been utilized as a fluorescent stain. It is membrane-permeable and selectively binds to adenine-thymine regions in the minor groove of B-form DNA with a high binding constant, making it a useful stain for DNA, chromosomes, and nuclei . It has been shown that the fluorescence is relatively insensitive to the presence of various solution reagents (i.e., BSA, RNA, Tris, sucrose, TEA/EDTA, Triton X-100, and SDS). Furthermore, the solution ionic strength and pH has been shown to be independent of the Stokes’ shift of bisBenzimide H 33258 . The properties of this dye make it useful in several applications such as detecting mycoplasma contamination in tissue culture , indicating early onset of apoptosis, sorting living cells based on DNA content, and acting as a probe for DNA-Drug interactions. Utilized in the design and development of a fluorimetric assay to quantitate cell numbers in microtiter plates. The methodology of the assay is based on Hoechst 33258 binding to DNA and is sensitive for a thousand cells. One application of this is assay is the normalization of ELISA data. The illumination of Hoechst 33253 coupled with Merocyanine 540 can be utilized as an effective method to eradicate bacterial infections (i.e., A. laidlawii contaminated corneal endothelial cell cultures) . BisBenzimide H 33258 is commonly used in many established staining protocols (David M. Terrian, Cancer Cell Signaling – Methods and Protocols,
Springer, Volume 218 ). |
| Application: |
bisBenzimide H 33258 is useful for staining DNA, chromosomes and nuclei. bisBenzimide H 33258 may be used for fluorescence microscopy or flow cytometry.
Excitation max. = 346 nm
Emission max. = 460 nm |
| Biochem/physiol Actions: |
bisBenzimide H 33258 is a potent and selective Bcl-XL inhibitor. bisBenzimide H 33258 is a membrane-permeable, fluorescent DNA stain with low cytotoxicity that intercalate in A-T regions of DNA. |
| Other Notes: |
Useful reagent for cytogenetic studies; for the fluorescent staining of DNA in cells; simple two-step fluorimetric assay of DNA in isolated nuclei, cell extracts, or crude homogenates using Hoechst 33258 |
| Symbol |
 GHS07 |
| Signal word |
Warning |
| Hazard statements |
H302 - H315 - H319 |
| Precautionary statements |
P305 + P351 + P338 |
| Hazard Codes |
Xn |
| Risk Statements |
22-36/38 |
| Safety Statements |
26-36 |
| RIDADR |
NONH for all modes of transport |
| WGK Germany |
WGK 3 |
| Flash Point(F) |
Not applicable |
| Flash Point(C) |
Not applicable |
| Purity |
≥98.0% (HPLC) |
| Storage Temp. |
2-8°C |
| UNSPSC |
12171500 |
