Whole-cell lysates were loaded onto 4 to 20% gradient SDS-PAGE gels (Bio-Rad) and transferred onto nitrocellulose membranes. The membranes were blocked with Block-One blocking reagent (Nakarai Tesque) and then incubated with primary antibodies using Can Get Signal Solution 1 (TOYOBO) overnight at 4 °C. The membranes were then incubated with secondary antibodies conjugated with horseradish peroxidase using Can Get Signal Solution 2 (TOYOBO) for 60 min at room temperature. Chemiluminescence signals developed with Chemi-Lumi One Super (Nakarai Tesque) were visualized by a ChemiDoc Touch and quantitated using ImageLab software (Bio-Rad).
4 to 20 gradient sds page gels
Manufactured by Bio-Rad
4 to 20% gradient SDS-PAGE gels are laboratory equipment used for protein separation and analysis. They provide a gradient of polyacrylamide concentrations, allowing for the separation of a wide range of protein sizes in a single gel. These gels are commonly used in the field of biochemistry and molecular biology for various applications.
Automatically generated - may contain errors
Lab products found in correlation
Can get signal solution 1, by Toyobo (1 mentions)
Can get signal solution 2, by Toyobo (1 mentions)
Block one blocking reagent, by Nacalai Tesque (1 mentions)
Chemi lumi one super, by Nacalai Tesque (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Chemidoc touch, by Bio-Rad (1 mentions)
Ab92726, by Abcam (1 mentions)
Clusterin, by Santa Cruz Biotechnology (1 mentions)
Filamin a, by Santa Cruz Biotechnology (1 mentions)
Gm130, by Santa Cruz Biotechnology (1 mentions)
Tsg101, by Abcam (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
2 protocols using 4 to 20 gradient sds page gels
1
SDS-PAGE Western Blot Protocol
2
Comprehensive Protein Analysis via SDS-PAGE
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Lysed samples were loaded onto two 4 to 20% gradient SDS-PAGE gels (Bio-Rad) and transferred to nitrocellulose membranes (Bio-Rad) by using standard methods. Gels were blocked with 5% bovine serum albumin in PBS with 0.05% Tween 20 (PBST). Antibodies used for Western blotting included 1:500 Tspan8 (sc-292058, Santa Cruz Biotechnology Inc.), 1:1000 TSG101 (ab83, Abcam), 1:1000 CD9 (ab92726, Abcam), 1:1000 GM130 (sc-55591, Santa Cruz Biotechnology Inc.), 1:1000 HSP70 (sc-32239, Santa Cruz Biotechnology Inc.), 1:1000 galectin-3–binding protein (sc-74970, Santa Cruz Biotechnology Inc.), 1:1000 clusterin (sc-8354, Santa Cruz Biotechnology Inc.), 1:1000 filamin-A (sc-28284, Santa Cruz Biotechnology Inc.), 1:1000 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2118L, Cell Signaling Technology), 1:1000 Itsn2 (ab133854, Abcam), and 1:1000 CD49d (PA520595, Invitrogen).
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