The indicated cells were incubated with DMSO, 5 µM vemurafenib, 10 µM SHP099, or the combination of 5 µM vemurafenib and 10 µM SHP099 for 0 h, 6 h, 24 h and 48 h. The cells were then lysed in RIPA lysis solution containing protease inhibitors for 30 min, and the protein concentration was determined using BCA reagent. Equal amounts of protein were separated by SDS-PAGE and transferred to PVDF membranes and incubated overnight at 4 °C with primary antibodies. The next day, co-incubation with the secondary antibody of the corresponding species was carried out for 2 h. Chemiluminescence detection was performed using the ECL Protein Blotting Assay Kit (Human IgG) (Solarbio). The primary antibodies were used: anti-SHP2 (Cell Signaling Technology, CAT #3397), anti-p-SHP2 (Cell Signaling Technology, CAT #3751), anti-ERK (Cell Signaling Technology, CAT #4695), anti-p-ERK (Cell Signaling Technology, CAT #4370), anti-AKT (Cell Signaling Technology, CAT #4685), anti-p-AKT (Cell Signaling Technology, CAT #4060), anti-MEK (Cell Signaling Technology, CAT #4694), anti-p-MEK (Cell Signaling Technology, CAT #9127).
Ecl protein blotting assay kit human igg
Manufactured by Solarbio
The ECL Protein Blotting Assay Kit (Human IgG) is a laboratory equipment product designed for the detection and analysis of human immunoglobulin G (IgG) proteins. The kit provides the necessary components for performing an enhanced chemiluminescence (ECL) protein blotting assay.
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Anti p shp2, by Cell Signaling Technology (2 mentions)
Anti mek, by Cell Signaling Technology (2 mentions)
Anti p mek, by Cell Signaling Technology (2 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
2 protocols using ecl protein blotting assay kit human igg
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Regulation of MAPK and AKT Signaling
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Molecular Signaling Cascades in BRAF-Mutant Cells
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The indicated cells were incubated with DMSO, 5 µM vemurafenib, 10 µM SHP099, or the combination of 5 µM vemurafenib and 10 µM SHP099 for 0 h, 6 h, 24 h and 48 h. The cells were then lysed in RIPA lysis solution containing protease inhibitors for 30 min, and the protein concentration was determined using BCA reagent. Equal amounts of protein were separated by SDS-PAGE and transferred to PVDF membranes and incubated overnight at 4 °C with primary antibodies. The next day, co-incubation with the secondary antibody of the corresponding species was carried out for 2 h. Chemiluminescence detection was performed using the ECL Protein Blotting Assay Kit (Human IgG) (Solarbio). The primary antibodies were used: anti-SHP2 (Cell Signaling Technology, CAT #3397), anti-p-SHP2 (Cell Signaling Technology, CAT #3751), anti-ERK (Cell Signaling Technology, CAT #4695), anti-p-ERK (Cell Signaling Technology, CAT #4370), anti-AKT (Cell Signaling Technology, CAT #4685), anti-p-AKT (Cell Signaling Technology, CAT #4060), anti-MEK (Cell Signaling Technology, CAT #4694), anti-p-MEK (Cell Signaling Technology, CAT #9127), anti-GAPDH (Cell Signaling Technology, CAT #5174).
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