HUVECs were stimulated with 20 μg/ml of FlgE or FlgEM for desired time lengths or pretreated with 50 μg/ml of anti-ATP5B antibodies for 1 h. Total proteins were extracted and subjected to 12% SDS-PAGE resolution, followed by transferring onto nitrocellulose membrane (Beyotime, Shanghai, China) for immunoblot analysis. The primary antibodies used were as follows: anti-VE-cadherin antibodies (1 μg/ml, Abcam), anti-phospho-VE-cadherin (Tyr731) antibodies (1 μg/ml, Affbiotech, Shanghai, China), anti-occludin antibodies (1 μg/ml, Abcam), anti-claudin-1 antibodies (1 μg/ml, Abcam), and anti-GAPDH antibodies (1 μg/ml, ABsin, Shanghai, China). Primary probing was carried out overnight at 4°C and, after washing, was incubated with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG antibodies (1:2,000, Cell Signaling Technology, Beverly, MA). The blots were detected using Enhanced Chemiluminescence (ECL) Amersham Western Blotting Detection Reagents (GE Healthcare).
Anti gapdh antibodies
Manufactured by Absin
Sourced in China
Anti-GAPDH antibodies are laboratory reagents used for the detection and quantification of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a commonly used housekeeping gene and is involved in the glycolytic pathway. These antibodies can be used in various analytical techniques, such as Western blotting, immunohistochemistry, and ELISA, to study the expression and localization of GAPDH in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti occludin antibody, by Abcam (1 mentions)
Horseradish peroxidase hrp conjugated anti rabbit igg antibody, by Cell Signaling Technology (1 mentions)
Anti ve cadherin antibody, by Abcam (1 mentions)
Nitrocellulose membrane, by Beyotime (1 mentions)
Enhanced chemiluminescence, by Cell Signaling Technology (1 mentions)
Pbs solution, by Thermo Fisher Scientific (1 mentions)
Anti nlrp3, by Absin (1 mentions)
2 protocols using anti gapdh antibodies
1
Immunoblot Analysis of Endothelial Junctions
2
NLRP3 Inflammasome Protein Expression Analysis
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Mononuclear cells were washed thrice with PBS solution (Invitrogen; Thermo Fisher Scientific, Inc.) and lysed in ice-cold lysis buffer (Absin Bioscience Inc., Shanghai, China). Quantitative analysis of protein concentrations was performed using bicinchoninic acid kit (Shanghai Weimeng Biotechnology Co. Ltd., Shanghai, China). The extracted proteins were added to SDS-polyacrylamide gel electrophoresis (12.5% gel) and transferred onto a nitrocellulose membrane. After blocking with 5% skim milk powder in TBST (Tris-buffered saline with 0.1% Tween 20), the membranes were incubated overnight at 4°C with the following primary antibodies: anti-NLRP3, anti–caspase l (1:1000, rabbit, Absin Bioscience Inc., Shanghai, China), anti-ASC, and anti-GAPDH antibodies (1:1000, rabbit, Absin Bioscience Inc., Shanghai, China). After washing, blots were incubated with secondary antibodies conjugated to HRP. The signals on the membrane were detected with enhanced chemiluminescence (Cell Signaling Technology).
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