Ag019 1

Manufactured by Beyotime

Sourced in China

The AG019‐1 is a lab equipment product. It is a specialized device designed for use in laboratory settings. The core function of the AG019‐1 is to perform specific tasks related to the needs of laboratory work.

Automatically generated - may contain errors

Lab products found in correlation

4 protocols using ag019 1

Western Blot Analysis of CUGBP1 Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols

Heart tissues were homogenized and lysed in RIPA lysis buffer (Beyotime Bio Co, China). Proteins were separated by 12% SDS/PAGE, transferred to PVDF membranes (Millipore, USA), blocked and incubated with antibodies. Final detection was performed using enhanced chemiluminescence detection solutions 1 and 2 (1:1) (ECL; Millipore). The relative protein expression level was analyzed by densitometry using ImageJ software (Bethesda, Maryland, USA). Primary antibodies used in this study were as follows: anti-CUGBP1 (sc-20003, Santa Cruz Biotechnology, USA), anti-Xpress (R910-25, Thermo, USA), β-actin (AA128-1, Beyotime, China) or GAPDH (AG019-1, Beyotime, China) were detected as control.

Liu Y., Wang H., Zhang H., Wang J., Liu Q., Bi Y., Song S., Qiao X., Zhu K., Wu Y, & Ji G. (2022). CUGBP1, a crucial factor for heart regeneration in mice. Cell Death & Disease, 13(2), 120.

+ Open protocol

+ Expand

Protein Extraction and Antibody Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell lysates were prepared by extracting proteins with RIPA buffer. The membranes were blocked in 5% non-fat dried milk and incubated overnight at 4°C with the appropriate primary antibodies. The rabbit anti-human VASH2 monoclonal antibody utilized in the western blot analyses was kindly provided by Professor Sato. The antibodies against GAPDH (AG019-1, Beyotime, China), Vimentin (ab8069, Abcam, UK), E-cadherin (ab1416, Abcam), ZEB1 (ab124512, Abcam), ZEB2 (ab25837, Abcam), ABCC1 (ab84320, Abcam), MMP2 (ab86607, Abcam), Bcl-2 (#2870, Cell Signaling Technology, USA) and CXCR4 (ab2074, Abcam) are commercially available.

Xue X., Zhang Y., Zhi Q., Tu M., Xu Y., Sun J., Wei J., Lu Z., Miao Y, & Gao W. (2014). MiR200-upregulated Vasohibin 2 promotes the malignant transformation of tumors by inducing epithelial-mesenchymal transition in hepatocellular carcinoma. Cell Communication and Signaling : CCS, 12, 62.

+ Open protocol

+ Expand

Western Blotting of EMT and Stemness Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blotting was performed using standard protocols. Briefly, cells were sonicated in RIPA buffer, and 30 μg proteins was resolved on 12% polyacrylamide SDS gels and transferred to polyvinylidene difluoride (PVDF) membranes. The membranes were blocked with 5% BSA (bovine serum albumin) in TBS (tris‐buffered saline), incubated overnight at 4°C with the appropriate primary antibodies. The antibodies used included those against VASH2 (ab116640, Abcam, Cambridge, UK), GAPDH (AG019‐1, Beyotime, Shanghai, China), Vimentin (ab8069, Abcam), E‐cadherin (ab11512, Abcam), ZEB1 (ab124512, Abcam), ZEB2 (ab25837, Abcam), Bcl‐2 (ab32124, Abcam), SMO (ab38686,, Abcam), Gli‐1 (ab49314, Abcam), and Gli‐2 (ab167389, Abcam).

Zhang Y., Xue X., Zhao X., Qin L., Shen Y., Dou H., Sun J., Wang T, & Yang D. (2018). Vasohibin 2 promotes malignant behaviors of pancreatic cancer cells by inducing epithelial‐mesenchymal transition via Hedgehog signaling pathway. Cancer Medicine, 7(11), 5567-5576.

+ Open protocol

+ Expand

Protein extraction and Western blotting protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins were extracted from cells or liver tissues in the lysis buffer (Beyotime, Shanghai, China) consisting of protease inhibitor cocktail (1:100, TargetMol, MA, USA). The concentration of proteins was determined using BCA Protein Assay kit (Beyotime). The extracted proteins were separated by 10% SDS/PAGE and electro‐transferred to PVDF membranes (Millipore, Billerica, MA, USA). After blocking in 5% milk for 1 h, the membranes were probed with primary antibodies against SLC27A5 (1:1000, NBP2‐37412, Novus Biologicals, CO, USA), α‐SMA (1:1000, ER1003, Huabio, Hangzhou, China), COL1A1 (1:1000, WL0088, Wanleibio, Shenyang, China), COL3A1 (1:1000, 22734‐1‐AP, Proteintech, IL, USA), RUNX2 (1:1000, 20700‐1‐AP, Proteintech, IL, USA), EGR3 (1:500, sc‐390967, Santa Cruz Biotechnology, USA), β‐actin (1:3000, BL005B, Biosharp), or GAPDH (1:3000, AG019‐1, Beyotime, Shanghai, China) at 4 °C overnight. Membranes were then incubated with horseradish peroxidase‐conjugated secondary antibody (Abcam, Cambridge, UK). The staining was visualized using ClarityTM Western ECL Substrate (Bio‐Rad, CA, USA).

Wu K., Liu Y., Xia J., Liu J., Wang K., Liang H., Xu F., Liu D., Nie D., Tang X., Huang A., Chen C, & Tang N. (2023). Loss of SLC27A5 Activates Hepatic Stellate Cells and Promotes Liver Fibrosis via Unconjugated Cholic Acid. Advanced Science, 11(2), 2304408.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!