Total protein was collected using RIPA buffer (Thermo Scientific, CA, USA), followed by protein quantification using Pierce™ BCA Protein Quantification Kit (Thermo Scientific). Then, 20 μg protein was used for electrophoresis loading, transfer and blocking. After incubation with primary antibody and corresponding secondary antibody, the blot was developed using SuperSignal™ West Dura Persistence Substrate (Thermo Scientific). The primary antibody used in this study: anti-YTHDC2 (#ab271139, Abcam), anti-E-Cadherin (#14472, CST), anti-N-Cadherin (#13116, CST), anti-YAP (#14074, CST), and anti-Tubulin (#2148, CST).
Supersignal west dura persistence substrate
Manufactured by Thermo Fisher Scientific
SuperSignal™ West Dura Persistence Substrate is a chemiluminescent detection reagent designed for Western blot analysis. It provides a sensitive, stable signal that can persist for hours, allowing for longer exposure times and the detection of low-abundance proteins.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (4 mentions)
Pierce bca protein quantification kit, by Thermo Fisher Scientific (2 mentions)
Anti rock2, by Cell Signaling Technology (1 mentions)
Anti kiaa1429, by Cell Signaling Technology (1 mentions)
Beta actin, by CWBIO (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
2 protocols using supersignal west dura persistence substrate
1
Western Blot Analysis of EMT Markers
2
Protein Quantification and Western Blotting
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Total proteins were collected using RIPA buffer (Thermo Scientific, CA, USA) and then quantified for protein quantification by using Pierce™ BCA Protein Quantification Kit (Thermo Scientific) [15 (link)]. Protein (20 μg) was used for electrophoresis loading SDS-PAGE and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% skimmed milk and incubated with primary antibodies, including anti-KIAA1429 (Cell Signaling Technology, 1:1000, #88,358), anti-ROCK2 (Cell Signaling Technology, 1:1000, #47,012), and beta-actin (CWBio, Beijing, China). After incubation by primary antibodies or their corresponding secondary antibodies, blots were developed using SuperSignal West Dura Persistence Substrate (Thermo Scientific).
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