Nuclear lysates were prepared by using NE-PER Nuclear and Cytoplasmic extraction reagents (Thermo Scientific) according to the manufacturer’s instructions. The proteins were separated by 12% SDS-polyacrylamide gel electrophoresis and transferred onto polyvinylidene difluoride membranes. The membranes were placed in 5% dry milk blocking buffer for 1h on a shaker at room temperature. The membranes were then washed and incubated with unconjugated α-Eos (1 (link):1000) overnight at 4°C. On the next day, the membranes were washed, and incubated with secondary antibody (anti-rat IgG) for 3h at 4°C. The membranes were washed extensively and incubated in developing solution (GE Healthcare, Chalfont St. Giles, Buckinghamshire, UK) for 5 minutes before exposure. Expression of β-actin was used as loading control.
Developing solution
Manufactured by GE Healthcare
Sourced in United Kingdom
The Developing Solution is a chemical formulation used in the processing of medical imaging films. It plays a crucial role in the development process, enabling the visualization of diagnostic images captured on radiographic film.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using developing solution
1
Western Blot Analysis of Eosinophil Proteins
2
Western Blot Protein Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were lysed using RIPA lysis buffer and lysates were separated by 10-15% sodium dodecyl sulfatepolyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. The membranes were incubated overnight with primary antibodies, the detailed information of which is described in Supplemental Methods. After washing, the membranes were incubated for 1 h in horseradish peroxidase-conjugated secondary antibodies. The membranes were incubated in a developing solution (GE Healthcare) and signal was detected using Chemi-Doc System (Bio-rad)..
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!