Lung tissue protein was extracted by RIPA buffer (Beijing Solarbio Science & Technology Co., Ltd., China). After the quantification of proteins, they were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred on NC membranes. The membranes were blocked with 5% skim milk and then incubated with primary antibodies against the target proteins (Abcam, Cambridge, UK; Abmart Inc., Shanghai, China) at 4°C overnight. Then, the membranes were incubated with secondary antibodies (Beijing Emarbio Science & Technology Co., Ltd.; China; anti-rabbit, Thermo Fisher Scientific, Inc., USA) for 1 h. The membranes were treated with enhanced chemiluminescence detection reagent (Biosharp, Shanghai, China), and the protein bands were visualized using Biorad Chemidoc XRS+ ChemiLuminescence Imaging System (Bio-Rad, USA). The relative band density was determined by Image J (National Institutes of Health, Bethesda, MD, USA).
Enhanced chemiluminescence detection reagent
Manufactured by Biosharp
Sourced in China
Enhanced chemiluminescence detection reagent is a laboratory product that enables the detection of proteins or other biomolecules through chemiluminescence. It serves as a reagent for the amplification and visualization of chemiluminescent signals.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti phospho akt, by Cell Signaling Technology (2 mentions)
Mouse anti akt, by Beyotime (2 mentions)
Imagequant las 500, by GE Healthcare (2 mentions)
Mouse anti β actin, by Merck Group (2 mentions)
Biorad chemidoc xrs chemiluminescence imaging system, by Bio-Rad (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
Ripa lysis buffer, by Solarbio (1 mentions)
5 protocols using enhanced chemiluminescence detection reagent
1
Western Blot Analysis of Lung Proteins
2
Western Blotting for AKT Signaling
3
Western Blotting for AKT Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was conducted using standard techniques with primary antibodies: mouse-anti-AKT (Beyotime Biotechnology), rabbit anti-phospho-AKT (Cell Signaling), and mouse-anti-β-actin (Sigma). Signals were assessed by an enhanced chemiluminescence detection reagent (Biosharp, China) and quantitated by densitometry (ImageQuant Las 500, GE Healthcare).
4
Western Blot Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H1688 and H446 cells were washed twice with PBS and lysed in radioimmunoprecipitation (RIPA) buffer to extract the total proteins. The protein content was measured with a Bradford protein assay kit. Equal proteins amount were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene fluoride (PVDF) membrane for incubating with diluted primary antibodies (1:1000) at 4℃ overnight. Then, the membrane was twashed three times with TBST at room temperature and incubated with the appropriate horseradish peroxidase (HRP)-linked secondary antibody for 1 h at room temperature. Proteins were visualized using enhanced chemiluminescence detection reagents (Biosharp, Guangzhou, China).
5
Western Blot Analysis of ARPE-19 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ARPE-19 in each group of animal organ tissue was lysed using RIPA lysis buffer (Solarbio Biotechnology, Beijing, China). Equal amounts of proteins were separated with SDS-polyacrylamide gels and then transblotted onto a PVDF membrane. The membranes were incubated with 5% dried nonfat milk buffer for 2 h to prevent nonspecific binding. They were then incubated with the relevant primary antibodies and with a secondary antibody. The protein bands in the membranes were detected by enhanced chemiluminescence detection reagents (Biosharp, Anhui, China) and analyzed with Image J.
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!