The total protein was extracted from the retina of 15 mice at P17 from each group by radio-immunoprecipitation assay (RIPA), then the protein was denatured at 100°C for 5 minutes and separated according to the instructions of 10% rapid gel preparation kit (Yamei, Shanghai, China). A voltage of 80 V was used for the electrophoresis and 200 mA was used for the PVDF membrane transfer, then 5% skim milk was used to block nonspecific protein. The blots were probed overnight at 4°C with the following primary antibodies PI3K, Akt, p-PI3K, p-AKT, VEGF and β-actin (all at dilutions 1:2000, Santa Cruz Biotechnology); IL-1β, IL-6, and TNF-α (all at 1:1000, ImmunoWay Biotechnology Company, Plano, TX, USA) Next, the blots were incubated with horseradish peroxidase-conjugated anti-rabbit secondary antibodies (1:2000, Cat# 65-6120, Molecular Probes) for 2 hours at room temperature, and were visualized using enhanced chemiluminescence (Millipore, Billerica, MA, USA). Finally, the optical density of protein bands was detected using GE AI680 imaging system (GE, Boston, MA, USA). The optical density ratio of target protein to β-actin was regarded as the relative protein expression.
Ge ai680 imaging system
Manufactured by GE Healthcare
Sourced in United States
The GE AI680 imaging system is a laboratory equipment designed for high-quality imaging. It utilizes advanced imaging technology to capture detailed visual data. The core function of the GE AI680 is to provide accurate and reliable imaging capabilities for various applications.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence, by Merck Group (1 mentions)
Vascular endothelial growth factor (vegf), by Santa Cruz Biotechnology (1 mentions)
P akt, by Santa Cruz Biotechnology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
P pi3k, by Santa Cruz Biotechnology (1 mentions)
Interleukin 6 (il 6), by Immunoway (1 mentions)
Tnf α, by Immunoway (1 mentions)
Il 1β, by Immunoway (1 mentions)
Hrp conjugated goat anti mouse igg, by Cell Signaling Technology (1 mentions)
Nitrocellulose blotting membrane, by GE Healthcare (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Tank binding kinase 1 tbk 1, by Cell Signaling Technology (1 mentions)
Stat1, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg, by Cell Signaling Technology (1 mentions)
2 protocols using ge ai680 imaging system
1
Retinal Protein Expression Analysis
2
Western Blot Analysis of PRRSV-Induced Immune Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells of the PRRSV-NI group and the PRRSV-ICs group at 6 hpi were lysed by the cell lysis buffer containing 1 mM of PMSF (Beyotime, Shanghai, China), and, respectively, mixed with 5 × SDS-PAGE loading buffer. The samples were separated by 8% SDS-PAGE and were then transferred to the Nitrocellulose Blotting Membrane (GE Healthcare, Boston, MA, USA). The membranes were blocked with 5% BSA at room temperature for 2 h and were then incubated overnight at 4 °C with the primary antibodies, including the TANK-binding kinase 1 (TBK-1) (Cell Signaling Technology, Boston, MA, USA), signal transducer and the activator of transcription 1 (STAT-1) (Cell Signaling Technology, Boston, MA, USA), signal transducer and activator of transcription 2 (STAT-2) (Cell Signaling Technology, Boston, MA, USA) and Myxovirus resistance protein 1 (Mx1) (Santa Cruz, Dallas, TX, USA). After being washed three times with PBST, the membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (Cell Signaling Technology, Boston, MA, USA) or HRP-conjugated goat anti-mouse IgG (Cell Signaling Technology, Boston, MA, USA) at room temperature for 1 h. Finally, the signals were detected using the GE AI680 imaging system (GE Healthcare, Boston, MA, USA).
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!