Polyvinylidene di uoride transfer membranes

The isolated hippocampal samples were used for western blot (n = 4 in each group) to determine the synthesis of in ammasome proteins. The samples were lysed via a lysis buffer (RIPA) and centrifuged. Total Protein Kit, Micro (Sigma, USA) was used to detect the total protein concentration. After protein denaturation, 5 μg of protein was loaded on 10% SDS-PAGE, and separated proteins were put on polyvinylidene di uoride transfer membranes (Sigma, USA) and then incubated for one hour with speci c primary antibodies (Novus Biologicals, USA). Then, the procedure was followed by incubation of Membranes for one hour with anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies (Abcam, Germany). Protein bands were detected by the luminescent substrate solution (Sigma, USA). Finally, to quantify the speci c bands, ImageJ software (NIH, USA) was used. GAPDH (Thermoscienti c, USA) was used for normalization [33] .

Using western blot technique, the isolated hippocampal samples were used (n = 4 in each group) to determine the synthesis of in ammasome proteins. The samples were lysed via a lysis buffer (RIPA) and centrifuged. Total Protein Kit, Micro (Sigma, USA) was used to detect the total protein concentration. After protein denaturation, 5 μg of protein were loaded on 10% SDS-PAGE, and separated proteins were put on polyvinylidene di uoride transfer membranes (Sigma, USA) and then incubated for one hour with speci c primary antibodies (Novus Biologicals, USA). Then, the procedure was followed by incubation of membranes for one hour with anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibodies (Abcam, Germany). Protein bands were detected by the luminescent substrate solution (Sigma, USA). Finally, to quantify the speci c bands, ImageJ software (NIH, USA) was used. GAPDH (Thermoscienti c, USA) was used for normalization [33] .