Whole protein was extracted by lysis buffer and denatured through heating at 100 °C for 5 min. Protein concentration in the supernatant was quantified by a BCA Protein Assay Reagent kit (Pro.23228, Thermo Fisher Scientific, USA). Dodecyl sulfate-polyacrylamide electrophoresis was conducted with 20 μg total protein loaded in each well. After electrophoresis, separated proteins were transferred to a nitrocellulose membrane (Millipore, Italy) in buffer. Following primary antibody (MUC1, Proteintech, 19976-1-AP, China; XPNPEP2, GeneTex, GTX109995, Irvine, CA, USA) and secondary antibody (Proteintech, 10230-1-AP, China) incubation, target protein bands were detected by an enhanced chemiluminescence kit (NCI5079, Thermo Fisher).
Xpnpep2
Manufactured by GeneTex
Sourced in China, United States
XPNPEP2 is a laboratory product that functions as a dipeptidyl aminopeptidase. It catalyzes the removal of N-terminal dipeptides from polypeptides with a broad specificity.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay reagent kit, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Merck Group (1 mentions)
Nci5079, by Thermo Fisher Scientific (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
Image lab 4, by Bio-Rad (1 mentions)
Enhanced chemiluminescence system, by Thermo Fisher Scientific (1 mentions)
Gapdh ab9485, by Abcam (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Snail1, by Cell Signaling Technology (1 mentions)
2 protocols using xpnpep2
1
Protein Extraction and Western Blotting
2
Quantitative Western Blot Analysis of Protein Expression
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Total cell lysates were prepared in radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime, Shanghai, China) supplemented with a protease inhibitor cocktail (Roche, Mannheirn, Germany). Protein concentrations were determined using a bicinchoninic acid (BCA) assay (Thermo Fisher Scientific). Total lysates (40 µg per sample) were subjected to sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting with primary antibodies against the following proteins: XPNPEP2 (GTX109995, 1:1000; GeneTex, Irvine, CA, USA), glyceraldehyde 3-phosphate dehydrogenase (GAPDH; ab9485, 1:2000; Abcam, Cambridge, MA, USA), E-cadherin (#3195, 1:2000; Cell Signaling Technology), Snail1 (#3879, 1:2000; Cell Signaling Technology), and vimentin (#5741, 1:2000; Cell Signaling Technology). After the membranes were washed, they were incubated with the corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the proteins were detected using an enhanced chemiluminescence system (Pierce, Thermo Fisher Scientific). The statistical data of the protein levels in the western blot were analyzed using the analysis tool in Image Lab 4.1 (Bio-Rad, Hercules, California, USA). We selected the weakest protein band in each group as the reference band.
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