Hoechst 34580 (Synonyms: HOE 34580;Proamine;Hoechst-34580;Hoechst34580) |
| Catalog No.GC17657 |
The nucleic acid stain Hoechst 34580 (Ex/Em: 392/440 nm) is frequently utilized as a cell-permeable nuclear counterstain that emits a blue fluorescence upon binding to dsDNA.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 23555-00-2
Sample solution is provided at 25 µL, 10mM.
;)
_1-7.$$$37316672@70.jpg');)
;)
;)
_273-287.$$$36806353@46.jpg');)
_1-16.$$$37156877@44.jpg');)
;)
;)
;)
_1124-1138.$$$35908550@30.jpg');)
_766-780.$$$37100057@30.jpg');)
_418-432.$$$36893736@30.jpg');)
_240-255.$$$35108512@29.jpg');)
_533-545.$$$36543525@28.jpg');)
_264-276.$$$36600053@22.jpg');)
_2214998.$$$@0.jpg');)
The nucleic acid stain Hoechst 34580 (Ex/Em: 392/440 nm) is frequently utilized as a cell-permeable nuclear counterstain that emits a blue fluorescence upon binding to dsDNA. Hoechst 34580 is commonly employed in various studies related to cell counting, cell cycle analysis, and cell replication. It is particularly useful in identifying condensed nuclei in apoptotic cells, as well as in combination with BrdU staining for cell-cycle studies.
Hoechst dyes are also useful for monitoring cell viability by tracking changes in their emission spectra. As minor groove-binding DNA stains with AT selectivity, the Hoechst dyes are able to bind to all nucleic acids, but they show a greater fluorescence enhancement for AT-rich double-stranded DNA strands compared to GC-rich strands [1]. This property has been exploited to identify Q-bands in chromosomes, which are regions rich in AT base pairs that fluoresce brightly when stained with the quinacrine dye [2].
Fig. Fluorescence excitation and emission spectra of Hoechst 34580 bound to DNA
References:
[1]. Portugal J, Waring MJ. Assignment of DNA binding sites for 4′, 6-diamidine-2-phenylindole and bisbenzimide (Hoechst 33258). A comparative footprinting study. Biochimica et Biophysica Acta (BBA)-Gene Structure and Expression. 1988 Feb 28;949(2):158-68.
[2]. Weisblum B, Haenssler E. Fluorometric properties of the bibenzimidazole derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma. 1974 Sep;46(3):255-60.
1. Preparing Hoechst 34580 dye stock solution
1.1 Prepare the Hoechst 34580 dye stock solution by dissolving the contents of one vial (100 mg) in 10 mL of DMSO to create a 10 mg/mL (22.1 mM) solution.
Note: Hoechst dye has poor solubility, so sonicate as necessary to dissolve. The 10 mg/mL Hoechst stock solution may be stored at 2–6°C for up to 1-3 months or at ≤ –20°C for longer periods and should be away from light always.
2. Labeling cells
2.1 Culture cells in an appropriate medium and vessel for fluorescence microscopy.
2.2 Prepare the Hoechst staining solution by diluting the Hoechst stock solution 1:1,000 in PBS.
2.3 Remove the medium in the cell plates.
2.4 Add sufficient staining solution to cover the cells.
2.5 Incubate for 5–10 minutes, protected from light.
2.6 Optional: You may image directly in the staining solution, if you wish.
2.7 Remove the staining solution.
2.8 Wash the cells 3 times in PBS, 5 mins each time.
2.9 Image the cells, Excitation/Emission (nm): 392/440.
This protocol only provides a guideline, and should be modified according to your specific needs.
| Cas No. | 23555-00-2 | SDF | |
| Synonyms | HOE 34580;Proamine;Hoechst-34580;Hoechst34580 | ||
| Chemical Name | N,N-dimethyl-4-(6-(4-methylpiperazin-1-yl)-1H,3'H-[2,5'-bibenzo[d]imidazol]-2'-yl)aniline | ||
| Canonical SMILES | CN1CCN(C2=CC3=C(N=C(C4=CC5=C(N=C(C6=CC=C(N(C)C)C=C6)N5)C=C4)N3)C=C2)CC1 | ||
| Formula | C27H29N7 | M.Wt | 451.57 |
| Solubility | ≥ 6.46 mg/mL in DMSO, ≥ 15.87 mg/mL in Water with ultrasonic and warming | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.2145 mL | 11.0725 mL | 22.145 mL |
| 5 mM | 0.4429 mL | 2.2145 mL | 4.429 mL |
| 10 mM | 0.2214 mL | 1.1072 mL | 2.2145 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
Required fields are marked with *