Polyacrylamide Gel Electrophoresis:
Membrane:
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Western Blot:
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
SDS Page:Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Saline:Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Incubation:Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Control:Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo ( Bubalus bubalis )
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
Article Title: Novel targets identified by integrated proteomic and phosphoproteomic analysis in spermatogenesis of swamp buffalo (Bubalus bubalis).
Article Snippet: .. Equal amounts of proteins were separated by SDS-PAGE and transferred onto a polyvinylidene fluoride (PVDF) membrane using a semidry western blotting system (Bio-Rad).The membrane was blocked with 5% nonfat milk in Tris Buffered Saline Tween (TBST) solution for 2 h at room temperature and then incubated overnight at 4 °C with primary antibody against ODF1 (ab197029, Abcam, Cambridge, MA, USA), NUCKS1 (bs-7917R, Bioss Biotechnology Inc., Beijing, China), LMNA (phospho S392) (ab58528, Abcam), PSMA3 (phospho S250) (bs-9353R, Bioss Biotechnology Inc.), and β-actin (loading control) (bs-0061R, Bioss Biotechnology Inc.) at a dilution of 1:1,000. .. Membranes were washed with TBST buffer three times and incubated with horseradish peroxidase-conjugated secondary antibodies (CWBIO, Beijing, China) in TBST buffer for 1.5 h at room temperature.
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