Nuclear factor of activated T cell 5 (NFAT5) expression was assessed using the western blot assay, as previously described (21 (link)). Briefly, CTX-TNA2 cells were harvested and lysed in RIPA buffer (Beyotime, Shanghai, China). The mixture supernatants were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 10% gels and were then transferred onto polyvinylidene fluoride membranes. The membranes were incubated overnight with NFAT5 monoclonal antibody (dilution, 1:1000, ab137407, Abcam, San Diego, CA, USA) and GAPDH loading control monoclonal antibody (dilution, 1:10,000, MA5-15738, Ebioscience, San Diego, CA, USA). The bound antibodies were detected using horseradish peroxidase-conjugated goat anti-mouse/rabbit IgG secondary antibody (dilution, 1:10,000, G-21040/21234, Ebioscience) and visualized using the enhanced chemiluminescent reagent (PerkinElmer Life Sciences, MA, USA).
Ab137407
Manufactured by Abcam
Sourced in United States
Ab137407 is an immunohistochemistry antibody. It is designed for the detection of target protein in tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Beyotime (1 mentions)
Enhanced chemiluminescent reagent, by PerkinElmer (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Pvdf membrane, by Beyotime (1 mentions)
Ecl kit, by Solarbio (1 mentions)
Ab134175, by Abcam (1 mentions)
Ab97779, by Abcam (1 mentions)
Ab97051, by Abcam (1 mentions)
2 protocols using ab137407
1
Quantifying NFAT5 Expression by Western Blot
2
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein of H1299 and A549 cells was extracted with radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime). Proteins were subsequently separated by electrophoresis using SDS‐PAGE gel and then transferred onto polyvinylidene difluoride (PVDF) membrane (Beyotime). The membrane was maintained overnight with primary antibodies including cyclin D1 (1:10 000, ab134175), MMP2 (1:1000, ab97779), NFAT5 (1:500, ab137407) or GAPDH (1:10 000, ab181602) that were purchased from Abcam. Afterwards, these membranes were incubated with secondary antibodies (ab97051, 1:5000, Abcam). The protein blots were visualized by using enhanced chemiluminescence (ECL) kits (Solarbio).
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