Total proteins were isolated from cell lysates using RIPA buffer containing protease inhibitors and phosphatase inhibitors (Thermo scientific, Rockford, IL). The protein extractions were then resolved on SDS-PAGE and transferred onto nitrocellular membranes (Millipore, Billerica, MA). After blocking in 5% skim milk, the membrane was incubated with primary antibodies overnight at 4°C. After washing, the horseradish peroxidase (HRP)-conjugated secondary antibodies were added and incubated for 1 h at room temperature. The blots were visualized using an enhanced chemiluminescence detection kit (Thermo scientific, Rockford, IL). Primary antibodies against TRIM35 and phosphorylated PKM2 (p-PKM2(Tyr105)) (Cell Signaling Technology, Danvers, MA) were used in Western blot analysis.
P pkm2 tyr105
Manufactured by Cell Signaling Technology
Sourced in United States
P-PKM2(Tyr105) is a laboratory reagent used for the detection and quantification of phosphorylated pyruvate kinase M2 (PKM2) protein. PKM2 is an enzyme involved in the glycolytic pathway and its phosphorylation at tyrosine 105 is a regulatory mechanism. This product can be used in various analytical techniques, such as Western blotting, to study the regulation and activity of PKM2 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor, by Thermo Fisher Scientific (1 mentions)
Phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescence detection kit, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Odyssey imaging system, by LI COR (1 mentions)
Lin28a, by Cell Signaling Technology (1 mentions)
Snap23, by Proteintech (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
C myc, by Cell Signaling Technology (1 mentions)
β actin, by Proteintech (1 mentions)
Calnexin, by Cell Signaling Technology (1 mentions)
Atp5a1, by ABclonal (1 mentions)
Tsg101, by Abcam (1 mentions)
2 protocols using p pkm2 tyr105
1
Isolation and Western Blot Analysis of TRIM35 and p-PKM2
2
Immunoblot Analysis of Cellular Proteins
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The cell and tissue lysates were extracted using RIPA lysis buffer containing protease and phosphatase inhibitor cocktail (NCM Biotech, Suzhou, China). Equivalent amounts of protein were electrophoresed on SDS-PAGE gels followed by transferring to PVDF membranes (Millipore, Billeria, MA). Antibodies against SNAP23 (Proteintech, China), TSG101, CD9 (Abcam, USA), Calnexin, Lin28a, Cyclin-D1, c-Myc, PKM2, p-PKM2 (Tyr105) (Cell Signaling Technology, USA) and ATP5A1, SDHA, CYTb, COX1 (ABclonal, Wuhan, China) were used for immunoblotted. β-Actin (Proteintech, China) served as the loading control. Protein bands were visualized and analyzed using the Odyssey Imaging System (LI-COR, USA) and Image J software.
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