Application Data
(NIH-3T3 cells were grown in 96 well plates, fixed with paraformaldehyde and permeabilized. Then incubate with DAPI solution for 5 minutes.)
DAPI Staining Solution IHC Related Reagent
DAPI Staining Solution
Applications
Immunohistochemistry, Flow Cytometry, Functional Assay, Immunofluorescence
Synonyms
DAPI Staining Solution; N/A; DAPI Staining Solution ihc related reagent
Form/Format
Liquid; 8 mM sodium phosphate, 2 mM potassium phosphate, 140 mM sodium chloride, 10mM potassium chloride; pH 7.4.
Concentration
1µg/ml (varies by lot)
Applicable Applications for DAPI Staining Solution ihc related reagent
IHC (Immunohistochemistry), FCM/FACS (Flow Cytometry), IF (Immunofluorescence)
Material
DAPI dihydrochloride
Stability
When stored as directed, product should be stable for one year.
Molecular formula
C16H15N5 - 2 HCL
Excitation: DAPI
340 nm
Emission: DAPI
488nm
Excitation: DAPI-DNA complexes
360nm
Emission: DAPI-DNA complexes
460nm
Notes
| Type of DAPI | Molecular formula | Molecular weight | Catalog Number |
|---|---|---|---|
| DAPI dihydrochloride | C16H15N5 - 2 HCl | 350.25 | MBS176580 |
| DAPI dilactate | C16H15N5 - 2 C3H6O3 | 457.48 | Please Inquire |
Enzymatic Activity
DNase-, RNase-, Protease- free
Cell Permeability
Intact membrane permeant
Sub-Cellular Localization
Nucleus
Biomolecule Reactivity
Specific binding to AT-base pairs; intercalation into GC-base pairs
Fluorescence spectrum
White-blue fluorescence
Usage and Handling
- For DNA and chromosomes staining, In vitro laboratory use only.
- DAPI is a known mutagen and should be handled with care
Protocol
- Fixed cells and tissues: wash appropriately to remove fixative. If necessary, immunofluorescent staining can be performed first, then perform the DAPI staining. If there is no other staining, perform DAPI staining directly.
Adherent cells or tissue slices: add a sufficient volume of DAPI stain solution to completely cover the sample.
Suspension cells: Add DAPI stain solution to suspend cell pellets at 3:1. Vortex to obtain an even suspension. - Incubate for 5-10 minutes at RT(room temperature)
- Remove the stain solution.
- Wash with TBST, PBS or physiological saline for 2-3 times, 3-5 minutes each.
- Observe the sample under fluorescence microscope (excitation wavelength: 360nm, emission wavelength:460nm)
Product Note
- Use Boster Antifade Mounting Medium (Product No. MBS176571) to prevent rapid photobleaching of fluorescence within spectrum range
- Please wear the lab coat and disposable gloves to operate.
Chemical Information
| Chemical Name | 4',6-Diamidino-2-phenylindole dihydrochloride |
| IUPAC Name | 2-(4-Amidinophenyl)-1H-indole-6-carboxamidine dihydrochloride |
| Molecular Formula | C16H15N5-2HCl; C16H17Cl2N5 |
| Molecular Weight | 350.25 |
| CAS | 28718-90-3 |
| EC Number | 249-186-7 |
| Chemical Sources |
ChEMBL: CHEMBL48217; CHEMBL531243; SCHEMBL2464378; CHEMBL545547 CHEBI: CHEBI:51231 ChemSpider: 2848; 123458; 140783 PubChem: 2954; 21924871; 67094309; 160166 |
Preparation and Storage
Store at -20°C, protect from light.
Application Data
(NIH-3T3 cells were grown in 96 well plates, fixed with paraformaldehyde and permeabilized. Then incubate with DAPI solution for 5 minutes.)
Application Data
(NIH-3T3 cells were grown in 96 well plates, fixed with paraformaldehyde and permeabilized. Then incubate with DAPI solution for 5 minutes.)
Related Product Information for DAPI Staining Solution ihc related reagent
DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye which can bind DNA strands robustly, the fluorescence being detected by fluorescence microscope. DAPI can dye both live and fixed cells as it can cross intact membrane, with higher efficiency in fixed cells. The molecular formula is C16H15N5·2HCl with 350.25 g/mol molecular weight, CAS Number 28718-90-3. DAPI could pass through the cell and nucleic membranes and bind the double-strand DNA in the nucleus, producing 20 times stronger fluorescence than itself. The efficiency detected by fluorescence microscope is very high (almost 100%), having no side effects for the live cells. The sensitivity for double stranded DNA DAPI staining is many times larger comparing to ethidium bromide (EB). DAPI staining is usually used in cell death detection, as it enters more effectively and generates stronger fluorescence in dead cells. After staining with DAPI, detect with fluorescence microscope or flow cytometry. Blue fluorescent cell would be seen under the microscope after staining. The largest excitation wavelength for DAPI is 340nm (ultraviolet), and the largest emission wavelength is 488nm (blue). When DAPI binds with double-strand DNA, the largest excitation wavelength is 360nm, while the largest emission wavelength becomes 460nm. DAPI's blue emission makes it suitable for combined assays where the fluorescence ranges of DAPI and other IHC-employed fluorescent molecules like green-fluorescent fluorescein and GFP, or red- fluorescent stains like Texas Red, are completely distinctive.
Product Categories/Family for DAPI Staining Solution ihc related reagent
Similar Products
Product Notes
The DAPI Staining Solution (Catalog #AAA45595) is an IHC Related Reagent and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's DAPI Staining Solution can be used in a range of immunoassay formats including, but not limited to, IHC (Immunohistochemistry), FCM/FACS (Flow Cytometry), IF (Immunofluorescence). Researchers should empirically determine the suitability of the DAPI Staining Solution for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "DAPI Staining Solution, IHC Related Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
If you are ready to order, navigate to Shopping Cart and get ready to checkout.