The detailed procedure was described in our previous study [41 (link)]. Firstly, cells were scraped from the 6-well culture plates for centrifugal collection at 300×g for 5 min at 4 °C. Cell-derived protein lysate was harvested by RIPA lysate (Beyotime, China). Ten micrograms of protein was fractionated by 10% SDS/PAGE, transferred to polyvinylidene difluoride membranes, and then incubated with the primary antibody at 4 °C overnight. After incubating with a secondary antibody for 45 min, the ECL system was used to detect the intensity of protein expression [42 (link), 43 (link)]. Primary antibodies were SLC7A11/xCT (1:1000, proteintech, China), ALDH1A1 (1:1000, proteintech, China) and Oct3/4 (1:1000, Wanlei, China), CD44 (1:1000, Wanlei, China), and GAPDH (1:5000, YI FEI XUE BIOTECHNOLOGY, Nanjing, China) was used as an internal reference.
Oct3 4
Manufactured by Wanlei
Sourced in China
The Oct3/4 is a laboratory equipment used for the detection and analysis of the Oct3/4 protein, a key transcription factor involved in the maintenance of pluripotency in stem cells. The device utilizes advanced techniques to accurately measure the expression levels of the Oct3/4 protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Aldh1a1, by Proteintech (2 mentions)
Slc7a11 xct, by Proteintech (1 mentions)
Ripa lysate, by Beyotime (1 mentions)
Minute total protein extraction kit, by Invent Biotechnologies (1 mentions)
Anti p erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Pakt s473, by Proteintech (1 mentions)
Erk1 2, by Wanlei (1 mentions)
β actin, by Beyotime (1 mentions)
Gapdh, by Beyotime (1 mentions)
2 protocols using oct3 4
1
Western Blot Analysis of Stem Cell Markers
2
Protein Extraction and Western Blotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Detailed procedure was described previously.54 (link) Protein in fresh tissues was extracted using Minute Total Protein Extraction Kit (Invent). Antibodies against the following proteins were used: YY1 (1:1,000, Wanleibio), OCT3/4 (1:1,000, Wanleibio), ALDH1A1 (1:1,000, Proteintech), p-AKT-s473 (1:1,000, Proteintech), AKT (1:1,000, Wanleibio), anti-p-ERK1/2 (Thr202/Tyr204; 1:1,000, Cell Signaling Technology), ERK1/2 (1:1,000, Wanleibio), GAPDH (glyceraldehyde-3-phosphate dehydrogenase 1:2,000, Beyotime), and β-actin (1:5,000, Beyotime). Protein expression levels were quantified by density analysis using Quantity One Software and normalized to β-actin or GAPDH.
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