The level of total protein in the supernatant of the NSCLC cell lysis was determined by the BCA (bicinchoninic acid) Protein Assay Kit (Thermo Fisher Scientific, China). The samples were boiled for 10 min at 95°C. An aliquot of 30 mg of protein was separated using 10% SDS–PAGE gel, followed by PVDF membrane transfer, blocking for 1 h in 5% skim milk, and incubation with separate antibodies (ABCAm, Inc., USA)—1:1000 diluted antibody Ab127548 (anti-RNF180), 1:500 diluted antibody Ab39688 (anti-c-Myc), 1:5000 diluted antibody Ab227198 (anti-HK-2), 1:1000 diluted Ab101562 antibody (anti-LDHA), and 1:2000 diluted anti-GAPDH antibody (#5174, Cell Signaling Technology)—overnight at 4°C. Thereafter, the membranes were incubated with horseradish peroxidase secondary antibodies (A0208 (1:200), A0181(1:200), and A0216(1:200); Beyotime Biotechnology) at room temperature for 1 h. The ECL plus substrate (GE Healthcare, USA) with the LAS-400 Image Analyzer (FujiFilm Medical Systems, USA) was used for the detection of the horseradish peroxidase signal.
Ab127548
Manufactured by Cell Signaling Technology
Sourced in United States
Ab127548 is a primary antibody that recognizes a specific target protein. It is designed for use in various laboratory techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry. The antibody is produced and quality-tested by Cell Signaling Technology.
Automatically generated - may contain errors
Lab products found in correlation
Ab227198, by Cell Signaling Technology (2 mentions)
A0216, by Beyotime (2 mentions)
Ecl plus substrate, by GE Healthcare (2 mentions)
A0208, by Beyotime (2 mentions)
Las 400 image analyzer, by Fujifilm (2 mentions)
Ab101562, by Cell Signaling Technology (1 mentions)
Protein assay kit, by Thermo Fisher Scientific (1 mentions)
A0181, by Beyotime (1 mentions)
Anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Bicinchoninic acid (bca), by Thermo Fisher Scientific (1 mentions)
Antibody against gapdh, by Cell Signaling Technology (1 mentions)
2 protocols using ab127548
1
Quantification of Protein Expression in NSCLC
2
Protein Expression Analysis in NSCLC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The total protein level in the supernatant of NSCLC cell lysis was determined by BCA (bicinchoninic acid) protein assay kit (Thermo Fisher Scienti c, China). The samples were boiled for 10 min at a temperature of 95°C. An aliquot of 30 mg of protein was separated by SDS-PAGE (10% gel) followed by PVDF membranes transfer, 1 h blocking in 5% skim milk, and incubation with separate antibodies (Abcam, Inc., USA) of 1:1000 diluted antibody Ab127548 (anti-RNF180), 1:500 diluted antibody Ab39688 (anti-C-myc), 1:5000 diluted antibody Ab227198 (anti-HK-2), 1:1000 diluted Ab101562 antibody (anti-LDHA), and 1:2000 diluted antibody against GAPDH (#5174, Cell Signaling Technology) overnight at 4°C. Then the membranes were incubated with horseradish peroxidase secondary antibodies (A0208, A0181 and A0216, Beyotime Biotechnology) at room temperature for 1 h. The ECL plus substrate (GE Healthcare, USA) with LAS-400 image analyzer (FujiFilm Medical Systems, USA) was used for the detection of horseradish peroxidase signal.
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