DAPI Staining Solution (1mg/mL)

Figure 1: Neurofilament L and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-neurofilament L antibody (green). For protocol see #Protocol 1 in application notes below.

Figure 2: GFAP and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-GFAP antibody (green). For protocol see #Protocol 1 in application notes below.

Figure 3: MAP2 and DAPI co-staining in hippocampal cell culture.

DAPI is a DNA binding dye commonly used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio labels cell nuclei (blue) at 1µg/ml when co-stained with an anti-MAP2 antibody (green). For protocol see #Protocol 1 in application notes below.

Figure 4. Peak emission spectrum of oxazole yellow when excited at 360nm and excitation spectrum when recording at 460nm.

DAPI is a DNA binding dye with strong affinity for nucleic acids used to label cell nuclei in immunofluorescence experiments. DAPI from Hello Bio has an excitation peak at 361nm and an emission peak at 456nm. For protocol see #Protocol 2 in application notes below.

#Protocol 1: DAPI counterstaining of primary cultured neurones.

• Primary neurones were isolated and cultured from P2 rats and grown for three weeks before being fixed with 4% paraformaldehyde.
• Coverslips containing neuronal cell cultures were labelled for either MAP2, GFAP or Neurofilament L following standard immunohistochemical approaches.
• Coverslips were then submerged in 1µg/ml DAPI diluted in PBS for 1 minute.
• Following 2 x 5-minute washes in PBS coverslips were mounted and imaged with a fluorescent microscope.

#Protocol 2: Measurement of excitation and emission spectra of DAPI

•  DAPI was prepared at 10 ng/ml in PBS with 10 ng/ml empty plasmid DNA.
• Excitation and emission spectra were measured between 230nm and 700nm using a Tecan Infinite M200 PRO ELISA plate reader.

Aliquot DAPI ready made solutions and store at -20°C and avoid freeze thaw cycles. This compound is light sensitive; exposure to light may affect compound performance, we therefore recommend storing solutions in the dark and protecting from light.