Horseradish peroxidase hrp conjugated anti mouse immunoglobulin g igg

Proteins from human lens tissue and SRA01/04 cell lysates were separated by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to a polyvinylidene fluoride (PVDF) membrane. Primary mouse antibodies against caspase-3 (Abcam, UK), BCL2 associated X(Bax), B-cell lymphoma-2(Bcl-2), and GAPDH (Proteintech), as well as a secondary antibody, horseradish peroxidase (HRP)-conjugated anti-mouse immunoglobulin G (IgG; Sigma), were used. Band intensities were quantified by Quantity One software. The protein levels were normalized to the GAPDH level.

All recombinant L. plantarum was cultured in MRS broth supplemented with 10 μg/mL erythromycin at 30°C without shaking. Once the OD600 reached 0.3, 50 ng/mL sakacin P induction peptide (SppIP) was added to the medium to induce antigen expression. After 10 h of induction at 30°C, recombinant L. plantarum was collected and lysed by repeated freeze‒thaw cycles at −80°C to obtain the target proteins. The proteins were separated by SDS‒PAGE (10% acrylamide) and transferred to nitrocellulose membranes. The membrane was then blocked with 5% skim milk powder at room temperature (RT) for 1 h. The presence of profilin-FliC and profilin proteins was detected using anti-Flag Tag (1:1,000) (Sigma, Saint Louis) and anti-His Tag (1:1,000) (Sigma) monoclonal antibodies, respectively. The secondary antibody used was horseradish peroxidase (HRP)-conjugated anti-mouse immunoglobulin G (IgG) (Sigma).