Rabbit polyclonal antibody for pias1

MCEL (C₃H₈O₂), which has a purity of 99.5% and a CAS# of 109-84-4, is manufactured by BDH Laboratory Supplies, England. SACI (C₉H₁₀O₅ and 98% pure) was produced by AK Scientific, USA. The DNA primers for SOCS1, STAT1, and JAK1 were products of ShineGene Corporation, Shanghai, China. The rabbit polyclonal antibody for PIAS1 was produced by Cloud-Clone Corp, TX, USA. Rat-specific ELISA kits for NF-κB, iNOS, and COX-2 were produced by Elabscience Biotechnology Inc., Houston, TX, USA, while rat-specific IL-6 and TNF-α ELISA kits were produced by BioLegend, Beijing, China. All chemicals were purchased from certified outlets and were of analytical grades.

Immunohistochemical analyses of the testis and liver were carried out using the method described by Somade et al. [47 (link)]. The tissue-mounted slides were allowed to dry overnight, followed by deparaffinization and rehydration using xylene and stepwise concentrations of ethanol (100%, 95%, and 70%). After soaking the slides in 3% hydrogen peroxide, heat-induced antigen retrieval was performed in citrate buffer and cooled in cold water for 5 min. Endogenous peroxidase blocking was carried out for 5 min using 5% BSA. The slides were incubated at room temperature for 60 min with a rabbit polyclonal antibody for PIAS1 (Cloud-Clone Corp, TX, USA) diluted in 1% phosphate buffer saline. Washed slides were then incubated with secondary antibody for 30 min, and visualization of immune complexes was ensured using 0.05% DAB. The slides were counterstained with hematoxylin and viewed under a light microscope to determine the liver and testis-positive stained cells. To avoid any bias, the viewing and scoring of the PIAS1 positive cells were carried out by a pathologist in a neighboring university who was not informed about the entire study.