Total protein was extracted from HCC cells using RIPA lysis buffer and quantified using Pierce BCA Protein Assay kit according to the manufacturer's protocol (Thermo Fisher Scientific, Waltham, Massachusetts, USA). Protein extracts (10 μg) were separated by SDS/PAGE gel (Thermo Fisher Scientific) and then transferred to a nitrocellulose membrane (Thermo Fisher Scientific). After membrane was blocked with 5% non-fat dry milk dissolved in Tris-buffered saline with Tween-20 buffer for 1 h at room temperature, it was incubated at 4°C overnight with primary antibodies against RFX5 (1:1000; Proteintech), YWHAQ (1:10,000; Proteintech), p53 antibody (1:2000; Proteintech), and Bax antibody (1:1500; Proteintech) followed by incubation with HRP-conjugated secondary antibodies (1:5000; Proteintech) for 1 h at room temperature. The band were detected using a Western Lighting Plus-ECL (PerkinElmer, Waltham, Massachusetts, USA) and visualized on X-ray film.
Ywhaq
Manufactured by Proteintech
Sourced in United States
YWHAQ is a recombinant protein that functions as a molecular chaperone, playing a role in the regulation of a range of cellular processes. It is commonly used in biochemical and cell biology research applications.
Automatically generated - may contain errors
Lab products found in correlation
Sds page gel, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated secondary antibody, by Proteintech (1 mentions)
P53 antibody, by Proteintech (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Western lighting plus ecl, by PerkinElmer (1 mentions)
Bax antibody, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
2 protocols using ywhaq
1
Protein Extraction and Western Blot Analysis of HCC Cells
2
Western Blot Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were harvested and washed twice with PBS. They were then lysed with cold RIPA and centrifuged at 12,000 g at 4° C for 10 min. The supernatants were collected and assayed for protein by the BCA method. The tissue and cell homogenates were separated by SDS-PAGE (4-12% gradient) and transferred onto a PVDF membrane. The PVDF membranes were blocked by TBS-T with 5% milk for about 2 h. The PVDF membranes were then incubated with antibodies overnight at 4° C and incubated with secondary HRP-IgG antibodies (CST Biotechnolog, USA) for 2 h at RT.
The primary antibodies were as follows: SORBS2 (1:1000; Proteintech); YWHAQ (1:1000; Proteintech); GAPDH (1:1000; Proteintech); β-tubulin (1:1000; CST).
The primary antibodies were as follows: SORBS2 (1:1000; Proteintech); YWHAQ (1:1000; Proteintech); GAPDH (1:1000; Proteintech); β-tubulin (1:1000; CST).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!