Cells were lysed in Laemmli buffer and analyzed for total protein concentration as described (Karni et al. 2007 (link)). Fifty micrograms of total protein from each cell lysate was separated by SDS-PAGE and transferred onto a nitrocellulose membrane. The membranes were blocked with 5% milk and probed with specific antibodies. Bands were visualized using enhanced chemiluminescence detection. Primary antibodies are as follows: hnRNP A1 (mAb A1/55, 1:1000) (Allemand et al. 2005 (link)), hnRNP A2/B1 (1:1000, Santa Cruz), β-tubulin (1:1000, Sigma), β-catenin (1:2000, Sigma), β-actin (1:1000, Santa Cruz), T7 tag (1:5000, Novagen), phospho-MEK S217/221 (1:1000, Cell Signaling), total MEK (1:1000, Cell Signaling), phospho-ERK T202/Y204 (1:1000 Sigma), total ERK1/2 (1:1000, Cell Signaling), A-Raf (1:1000, Cell Signaling), and A-Raf short (1:500) (Rauch et al. 2011 (link)). Secondary antibodies are as follows: HRP-conjugated goat anti-mouse, goat anti-rabbit, donkey anti-goat IgG (H+L; 1:10,000 Jackson Laboratories).
Phospho mek s217 221
Manufactured by Cell Signaling Technology
Phospho-MEK S217/221 is a laboratory reagent used for the detection and quantification of MEK (Mitogen-Activated Protein Kinase Kinase) phosphorylated at Serine 217 and 221 residues. It is a specific antibody designed for use in techniques such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA) to study the activation of the MEK signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Total erk1 2, by Cell Signaling Technology (2 mentions)
Phospho erk t202 y204, by Merck Group (2 mentions)
β catenin, by Merck Group (2 mentions)
Total mek, by Cell Signaling Technology (2 mentions)
Hnrnpa1, by Santa Cruz Biotechnology (1 mentions)
Hnrnpa2 b1, by Santa Cruz Biotechnology (1 mentions)
Hrp conjugated goat anti mouse, by Jackson ImmunoResearch (1 mentions)
Donkey anti goat igg h l, by Jackson ImmunoResearch (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
β tubulin, by Merck Group (1 mentions)
Aldh1a1, by Santa Cruz Biotechnology (1 mentions)
P glycoprotein, by Abcam (1 mentions)
Hrp conjugated anti rabbit and mouse igg, by Bio-Rad (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Phospho stat3 y705, by Abcam (1 mentions)
Pbs tablet, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Gapdh, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Capmatinib, by Selleck Chemicals (1 mentions)
3 protocols using phospho mek s217 221
1
Western Blot Analysis of Protein Expression
2
Inhibition of Oncogenic Pathways
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Doxazosin mesylate, crizotinib, capmatinib, tepotinib, osimertinib and lazertinib were purchased from Selleckchem (Radnor, PA). Amivantamab was obtained from MedchemExpress (Monmouth Junction, NJ). Triton X-100, propidium iodide, PBS tablet, dimethyl sulfoxide (DMSO) and cOmplete™ protease inhibitor cocktail were obtained from Sigma-Aldrich (St. Louis, MO). The immunoblotting and immunostaining antibodies utilized were obtained as follows: c-MET, phospho-c-MET (Y1234/1235), EGFR, phospho-EGFR (Y1068), MEK, phospho-MEK (S217/221), AKT, phospho-AKT (S473), cleaved caspase-3 (Asp175), cleaved caspase-7 (Asp198), cleaved caspase-8 (Asp391), PARP, JAK2, CD44, OCT4 and SOX2 (Cell Signaling Technology, Beverly, MA); phospho-JAK2 (Y1007/1008), STAT3, phospho-STAT3 (Y705) and P-glycoprotein (Abcam, Cambridge, MA); survivin, cyclin D1 and ALDH1A1 (Santa Cruz Biotechnology, CA); GAPDH (Invitrogen, Carlsbad, CA). The secondary antibodies used were HRP-conjugated anti-rabbit and mouse IgG (Bio-Rad Laboratories Inc, CA) and Alexa Fluor-488 or -594 goat anti-mouse and rabbit IgG (Invitrogen).
3
Western Blot Analysis of Cell Signaling
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Cells were lysed in Laemmli buffer and analyzed for total protein concentration. 20 μg of total protein from each cell lysate was separated by SDS-PAGE and transferred on to a PVDF membrane. The membranes were blocked with 5% milk and probed with specific antibodies. Bands were visualized using enhanced chemiluminescence detection. Primary antibodies: β-Catenin (1:2000, Sigma), Phospho-MEK S217/221 (1:1000, Cell Signaling), total MEK (1:1000, Cell Signaling), phospho-ERK T202/Y204 (1:1000 Sigma), total ERK1/2 (1:1000, Cell Signaling), phospho-AKT (1:1000, Cell Signaling), AKT (1:1000, Cell Signaling), cleaved Caspase 3 (1:1000, Cell Signaling) SRSF1 (AK96 culture supernatant 1:300), CUGBP1 (1:1000, Santa Cruz Biotechnology), MBNL1 (1:250, Santa Cruz Biotechnology), SRSF3 (1:1000, Santa Cruz Biotechnology). Secondary antibodies: HRP-conjugated goat anti-mouse, goat anti-rabbit and donkey anti-goat IgG (H+L; 1:10000 Jackson Laboratories).
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