At 24 hrs after ROSC, the rats (n=8/group) were anesthetized with intraperitoneal injection of 2% pentobarbital sodium. Then, the brains were quickly removed. A quantity of 60 mg cerebral cortex tissue was taken, blood was washed with 1×PBS solution, and 600 μL1×PBS was added to make homogenate. The brain tissue homogenate was placed at −20°C refrigerator overnight. After repeated freezing and thawing for two times, the brain tissue homogenate was centrifuged at 5000 g at 4°C for 5 mins. The supernatant was then used for the measurement of brain ATP synthase (ATP5C1) according to Rat ATP synthase subunit ELISA kit (Wuhan Huamei Biological Engineering Co. Ltd., People's Republic of China). The absorbance (OD) value was determined by 450 nm wavelength multifunctional enzyme spectrometer (BIOTEK Synergy H1, USA), and the concentration of the sample ATP synthase (ATP5C1) was calculated. Tissue protein was quantified using the Coomassie brilliant blue reagent (Nanjing Jian Cheng Technology), and ATP synthase (ATP5C1) contents were calculated relative to the mass of protein (n=8 for each group).
Coomassie brilliant blue reagent
Manufactured by Nanjing Jiancheng
Sourced in China
Coomassie brilliant blue reagent is a protein staining solution used in biochemical analysis. It is a dye-based reagent that binds to proteins, allowing them to be visualized and quantified.
Automatically generated - may contain errors
3 protocols using coomassie brilliant blue reagent
1
Brain ATP Synthase Measurement in Rats
2
Quantifying Salt-Soluble Protein Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Salt soluble proteins (SSP) solubility was determined with a modification of the procedures of Frye et al. (1986) . Small sponges (2 cm×2 cm×2 cm) were added into tumbler together with the chop samples and marinade at the start of marination experiments. Then the sponge was removed immediately after experiments and pressed by a 50 mL volumetric syringe to get 4 g slurry. After vacuum treatment for 15 min, the slurry was added in 200 mL of 3% NaCl solution. After storing at 4°C for 24 h, the slurry solution was centrifuged for 15 min at 10,400×g at 4°C (Beckman Allegra 64R, Beckman-Coulter Company, Fullerton, CA, USA). The total protein content in the supernatant was determined by the Coomassie brilliant blue reagent (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) from the absorbance at 595 nm.
3
Measuring Brain ATP Levels in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 24 hrs after ROSC, the rats (n=8/group) were anesthetized with intraperitoneal injection of 2% pentobarbital sodium. Then, the brains were quickly removed. About 60 mg cerebral cortex tissues were collected and homogenized with 600 μL ATP detection cracking solution according to ATP detection kit (S0026, Beyotime Biotechnology Co. Ltd., Shanghai, People's Republic of China). The brain tissue homogenate was centrifuged at 12,000 g at 4°C for 5 mins. The supernatant was then used for the measurement of brain ATP contents according to ATP detection kit. Fluorescein in the ATP detection reagent uses the energy provided by ATP to produce fluorescein, which is directly proportional to the concentration of ATP, and uses a chemiluminescence detector (Luminometer) to detect fluorescence. Tissue protein was quantified using the Coomassie brilliant blue reagent (Nanjing Jian Cheng Technology, People's Republic of China), and contents of ATP were calculated relative to the mass of protein (n=8 for each group).
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!