Anti hdac1 h 51

Protein lysates were prepared from mouse CD4⁺ T cells using RIPA lysis buffer for Western blotting. After the quantification of protein concentrations using the BIO-RAD DC^TM protein assay, protein lysates were subjected to SDS-PAGE (10%). Blots were incubated with anti-HDAC1 (H-51) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-HDAC2 (H-54) (Santa Cruz Biotechnology), anti-HDAC3 (H-99) (Santa Cruz Biotechnology), anti-K_Ca3.1 (Alomone Labs, Jerusalem, Israel), and anti-ACTB (Medical & Biological Laboratories, Nagoya, Japan) antibodies, and were then incubated with anti-rabbit horseradish peroxidase-conjugated IgG (Merck, Darmstadt, Germany). An enhanced chemiluminescence detection system (Nacalai Tesque, Kyoto, Japan) was used to detect the bound antibody. The resulting images were analyzed using Amersham Imager 600 (GE Healthcare Japan, Tokyo, Japan). The light intensities of the band signals relative to that of the ACTB signal were calculated using ImageJ software (Ver. 1.42, NIH, Bethesda, MA, USA). In the summarized results, relative protein expression levels in the control were expressed as 1.0.