After harvesting cells, added cell lysis buffer (25 mM HEPES, pH 7.4; 150 mM NaCl; 1% NP-40; 0.25% sodium deoxycholate; 1 mM EGTA; 1 mM DTT; 50 µg/mL trypsin inhibitor; 1 mM PMSF; and 10 µg/mL aprotinin) into cells on ice for 30 minutes, centrifuged them at 12 000 rpm 4 °C for 10 minutes, took the supernatant, added SDS loading buffer and then boiled it at 100 °C for 5 minutes, performed western blotting analyses. Proteins of different groups were separated by electrophoresis in 12% sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane (GE, US), and rinsed three times (10 minutes each time) by TBS-T (10 mM Tris-Cl, pH 8.0, 150 mM NaCl, and 0.5% Tween 20). After 3 hours of incubation at room temperature with the rst antibodies (EGFP monoclonal antibody, Santa Cruz (sc-9996); survivin monoclonal antibody, Santa Cruz(sc-17779); β-actin monoclonal antibody, Santa Cruz (sc-81178); Bcl-2 polyclonal antibody, Boster (A00040-2), Ki67 polyclonal antibody, Boster (PB0065)), incubated with HRP-conjugated IgG (ThermoFisher) for 1 hour, then detected by the enhanced chemiluminescence system (ThermoFisher).
Survivin monoclonal antibody
Manufactured by Santa Cruz Biotechnology
The Survivin monoclonal antibody from Santa Cruz Biotechnology is a laboratory research tool used to detect the Survivin protein in various experimental applications. It is a highly specific and sensitive antibody that recognizes the Survivin protein, which is involved in the regulation of cell division and apoptosis.
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2 protocols using survivin monoclonal antibody
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Western Blot Analysis of Protein Expression
2
Western Blot Analysis of Protein Expression
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After harvesting cells, added cell lysis buffer (25 mM HEPES, pH 7.4; 150 mM NaCl; 1% NP-40; 0.25% sodium deoxycholate; 1 mM EGTA; 1 mM DTT; 50 µg/mL trypsin inhibitor; 1 mM PMSF; and 10 µg/mL aprotinin) into cells on ice for 30 minutes, centrifuged them at 12 000 rpm 4 °C for 10 minutes, took the supernatant, added SDS loading buffer and then boiled it at 100 °C for 5 minutes, performed western blotting analyses. Proteins of different groups were separated by electrophoresis in 12% sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane (GE, US), and rinsed three times (10 minutes each time) by TBS-T (10 mM Tris-Cl, pH 8.0, 150 mM NaCl, and 0.5% Tween 20). After 3 hours of incubation at room temperature with the rst antibodies (EGFP monoclonal antibody, Santa Cruz (sc-9996); survivin monoclonal antibody, Santa Cruz(sc-17779); β-actin monoclonal antibody, Santa Cruz (sc-81178); Bcl-2 polyclonal antibody, Boster (A00040-2), Ki67 polyclonal antibody, Boster (PB0065)), incubated with HRP-conjugated IgG (ThermoFisher) for 1 hour, then detected by the enhanced chemiluminescence system (ThermoFisher).
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