Total protein was isolated from the retinal samples and run on 10% polyacrylamide gels following a standard protocol. Equal amounts of protein resolved by polyacrylamide gels were immunoblotted using antibodies against β-tubulin (1:10000, RM2003; Beijing Ray Antibody Biotech, Beijing, China), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:10,000, RM2002; Beijing Ray Antibody Biotech), caspase-3 (1:1000, 9662; Cell Signaling Technology, Danvers, MA, USA), Bax (1:1000, 2772; Cell Signaling Technology), B-cell lymphoma-2 (Bcl-2; (1:1000, ab182858; Abcam, Cambridge, UK), Akt (1:1000, YT0177; ImmunoWay, Plano, TX, USA), phosphorylated Akt (p-Akt; 1:1000, YP0006; ImmunoWay), glycogen synthase kinase-3 beta (GSK3β; 1:1000, 12456; Cell Signaling Technology), phosphorylated GSK3β (p-GSK3β; 1:1000, 9323; Cell Signaling Technology), and β-catenin (1:1000, ab32572; Abcam). They were then detected with a chemiluminescence kit. Gray value analysis was conducted using Image J software.
Yt0177
Manufactured by Immunoway
Sourced in United Kingdom, United States
YT0177 is a laboratory equipment designed for general scientific applications. It serves as a basic tool for various experiments and research activities. The core function of YT0177 is to provide a platform for controlled and consistent sample handling and processing. The specific details and intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Yp0006, by Immunoway (2 mentions)
Ab182858, by Abcam (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
P akt, by Immunoway (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
Ab32572, by Abcam (1 mentions)
Polyvinyl difluoride membrane, by Merck Group (1 mentions)
Gb23303, by Wuhan Servicebio Technology (1 mentions)
Gb23301, by Wuhan Servicebio Technology (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
2 protocols using yt0177
1
Retinal Protein Expression Analysis
2
Western Blot Analysis of Protein Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells and tissues were lysed in RIPA buffer (Solarbio, Beijing, China), and proteins were separated by SDS-PAGE and transferred to polyvinyldi-fluoride membranes (Millipore, USA), and then the membranes were blocked with 5% blotting-grade milk for 1 h. the membranes were incubated overnight at 4°C with rabbit antibodies against CENPU (1:2,000, YN1585, Immunoway), rabbit antibodies against PI3K-p110α (1:1,000, YT3709, Immunoway), rabbit antibodies against AKT1 (1:1,000, YT0177, Immunoway), rabbit antibodies against pAKT Ser473 (1:1,000, YP0006, Immunoway), mouse antibodies against S6 (1:1,000, 2317S, Cell Signaling Technology), rabbit antibodies against pS6 Ser235/236 (1:1,000, YP0243, Immunoway), rabbit antibodies against NF-κB p65 (1:1,000, 8242S, Cell Signaling Technology), mouse antibodies against β-actin (1:1,000, 3700S, Cell Signaling Technology). The blots were then washed and incubated for 1 h at room temperature with peroxidase-conjugated anti-rabbit IgG and anti-mouse IgG secondary antibody (Servicebio, GB23301 and GB23303, respectively, 1:3,000). The concentration of protein fragments was measured using Image-Pro Plus Ver. 7.0 software. URL: https://www.mediacy.com/imageproplus .
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