Following treatment with 2 μg/ml ephrin-A1-Fc and/or inhibitors, cells were rinsed with ice cold PBS and processed according to the NE-PER cytoplasmic/nuclear subcellular fractionation kit (Pierce). Nuclear pellets were rinsed with ice cold PBS after extraction of the cytoplasmic fraction and fractions were diluted with 2X Laemmli sample buffer prior to analysis by immunoblotting. Immunoblotting was performed on cell lysates following separation on 10% polyacrylamide gels by SDS-PAGE and subsequent transfer to PVDF membranes. Membranes were blocked with 5% milk in tris-buffered saline containing tween-20, and then probed with rabbit anti-p65, rabbit anti-EphA4, rabbit anti-EphA2, rabbit anti-DSCR1, mouse anti-E-selectin (Santa Cruz); rabbit anti-phospho-p65, rabbit anti-NFAT1, rabbit anti-HDAC3, rabbit anti-β-tubulin, rabbit anti-GAPDH, or rabbit IκBα (Cell Signaling Technologies).
Rabbit anti nfat1
Manufactured by Cell Signaling Technology
Rabbit anti-NFAT1 is a primary antibody that specifically binds to the Nuclear Factor of Activated T-cells 1 (NFAT1) protein. NFAT1 is a transcription factor that plays a crucial role in the regulation of gene expression in various cell types, particularly in the immune system.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti p65, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti epha4, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti phospho p65, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti hdac3, by Cell Signaling Technology (1 mentions)
Rabbit anti β tubulin, by Cell Signaling Technology (1 mentions)
Rabbit anti epha2, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (1 mentions)
Mouse anti gst, by Cell Signaling Technology (1 mentions)
Rabbit anti laminb1, by Proteintech (1 mentions)
Mouse anti β actin, by Abbkine (1 mentions)
Ripa lysis solution, by Applygen (1 mentions)
2 protocols using rabbit anti nfat1
1
Ephrin-A1-Fc Signaling Pathway Analysis
2
Western Blot Analysis of Protein Targets
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The cells and tissues were lysed with an appropriate amount of RIPA lysis solution (Applygen, China) containing protease inhibitors, PMSF, and β-ME. The proteins were separated by SDS-PAGE and then transferred to PVDF membranes for primary antibodies incubation and secondary antibodies incubation. Mouse anti-GFP and mouse anti-β-actin were purchased from Abbkine. Rabbit anti-calcineurin A, mouse anti-GST, and rabbit anti-NFAT1 were purchased from Cell Signalling Technology. Rabbit anti-lamin B1 was provided by Proteintech. Goat anti-mouse IgG and goat anti-rabbit IgG secondary antibodies were purchased from SGB-BIO. After the secondary antibody incubation was completed, ECL luminescent solution (Tanon, China) was used for colour development. A Tanon instrument was used to obtain images, and ImageJ was used for image analysis.
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