ELISA kits were used to quantify the expression of TNF-α, COX-2, and p-NF-kB, according to manufacturer’s instructions. Then 2,500 µL of PBS was added to 50 mg of brain tissue and a small quantity of PMSF was also added to the solution as a protease inhibitor.56 (link) The mixture was first homogenized at 15,000 rpm and then centrifuged at 4,000x g for 10 minutes and the supernatant was collected. BCA method (Elabscience) was used to calculate the protein concentration in the supernatant of each group and an equal quantity of protein was loaded to quantify protein concentrations of TNF-α, COX-2, and p-NF-kB using an ELISA plate reader (BioTek EL×808). The protein concentration obtained (pg/mL) was then normalized to the total protein content (pg/mg total proteins).
Protocol detail
Quantifying Brain Inflammatory Markers
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.
Corresponding Organization :
Other organizations : Riphah International University, Shenzhen Children's Hospital, Peking University
Top 5 similar protocols
Variable analysis
independent variables
- ELISA kits used to quantify the expression of TNF-α, COX-2, and p-NF-kB
- Expression of TNF-α
- Expression of COX-2
- Expression of p-NF-kB
- 2,500 µL of PBS added to 50 mg of brain tissue
- PMSF added as a protease inhibitor
- Homogenized at 15,000 rpm
- Centrifuged at 4,000x g for 10 minutes
- Supernatant collected
- BCA method used to calculate protein concentration
- Equal quantity of protein loaded to quantify protein concentrations of TNF-α, COX-2, and p-NF-kB using an ELISA plate reader
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!