News from the Bench
Discover what’s going on at CST, receive our latest application notes and tips, read our science features, and learn about our products.
CST Antibody Performance Guarantee
To Purchase # 4083S
Find answers on our FAQs page.
PTM information and tools available.
Immunofluorescent analysis of HeLa cells using COX IV (3E11) Rabbit mAb #4850 (green), β-Catenin (L54E2) Mouse mAb (IF Preferred) #2677 (red) and DAPI (blue).Learn more about how we get our images
Gallery: DAPI #4083
DAPI is supplied as a lyophilized powder in 1 mg units. It can be used to examine cellular DNA in fluorescent microscopy and cytometry applications.Molecular Formula:
- Free dye excitation maximum = 340 nm
- Free dye emission maximum = 488 nm
- DNA complex excitation maximum = 364 nm
- DNA complex emission maximum = 454 nm
Directions for Use: DAPI is supplied as a lyophilized powder. For a 20 mg/ml stock, reconstitute the 1 mg in 50 µl deionized water or dimethylformimide. Please follow CST’s recommended IF and Flow protocols. For both applications, following secondary detection:
Immunofluorescence: Counterstain with DAPI as the final step in your staining procedure. Rinse samples twice in PBS for five min each. Dilute DAPI stock solution to a concentration between 1 - 0.1 µg/ml in PBS and incubate for 5 min at room temperature in the dark. Rinse samples once in PBS and then prepare for imaging. Examine immediately using appropriate excitation wavelength.
Alternatively, dilute DAPI stock solution to a concentration between 1 - 0.1 µg/ml in mounting media, apply to cells, and prepare for imaging. Examine immediately using appropriate excitation wavelength.
Flow Cytometry: Rinse samples once in Incubation Buffer. Dilute DAPI stock solution to a concentration between 1.60-0.0125 µg/ml in PBS and incubate for 15 min at room temperature in the dark before analyzing cells on flow cytometer.Storage: Storage lyophilized or in solution at 4°C, desiccated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 2-3 weeks to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) is a blue fluorescent DNA dye that targets double-stranded AT clusters in the DNA minor groove (1). One molecule of dye binds to each 3 base pairs of dsDNA and yields an approximate 20-fold fluorescent enhancement (2). The level of DAPI-DNA fluorescence is proportional to DNA content (3).
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc. XP® is a trademark of Cell Signaling Technology, Inc. Alexa Fluor® is a registered trademark of Life Technologies Corporation. DyLight™ is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.