Preparation of sections as previously described [21 (link)]. Membrane breaking was performed by PBS containing 0.1% triton-100 (PBS-T) at room temperature (RT) for 20 min. The sections were then blocked (RT, 1 h) using PBS-T containing 5% BSA. Sections were incubated (4°C, 16 h) using anti-glial fibrillary acidic protein (GFAP) (dilution 1:100, ab207165, Abcam, UK) and anti-S100β (dilution 1:100, ab52642, Abcam). After completion of incubation (RT, 1 h) with secondary antibodies (dilution 1 : 500, Alexa Fluor488, ab150077, Abcam; dilution 1 : 500, Alexa Fluor594, ab150080, Abcam), 4-6-diamidino-2-phenylindole (ZLI-9557, Zhongshanjinqiao Biotech, PRC) incubation (RT, 5 min) was performed. The staining was observed under a fluorescence microscope (BX-60, Olympus, JPN) and images were taken. The fluorescence intensity was analyzed using ImageJ 8.0, National Institutes of Health.
Protocol detail
Immunofluorescent Analysis of Astrocytes
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Antibodies
Dilution
Fluorescence
Fluorescence microscope
Glial fibrillary acidic protein
Membrane
S100β
Corresponding Organization : Ningxia Medical University
Top 5 similar protocols
Variable analysis
independent variables
- Membrane breaking method (PBS containing 0.1% triton-100 (PBS-T) at room temperature (RT) for 20 min)
- Fluorescence intensity of GFAP and S100β staining
- Incubation time of primary antibodies (4°C, 16 h)
- Incubation time of secondary antibodies (RT, 1 h)
- Incubation time of DAPI (RT, 5 min)
- Microscope used (BX-60, Olympus, JPN)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!