Cells were lysed in lysis buffer (25 mM Tris pH 8.0, 125 mM NaCl, 1 mM MgCl2, 1% Triton X-100, 1 mM sodium orthovanadate, 1 mM sodium fluoride, 1X protease inhibitor (Sigma-Aldrich), phosphatase inhibitor cocktails #2 and #3 (Sigma-Aldrich), and 1 mM PMSF). Cell lysates were resolved on a Nupage 4-12% Bis-Tris gradient gel (Life Technologies) and probed using antibodies recognising phospho- and total eIF4E1 (Cell Signaling #9741 and Santa Cruz sc-9976), phospho- and total ERK (#9101 and #9102, Cell Signaling), c-Myc (#9402), n-Myc (#9405), p-MNK1/2 (#2111), dicer (#3363), eIF4A (#2013) (Cell Signaling), GAPDH (ab8245), BTK (ab54219), YY1 (ab12132) (Abcam), eIF4E3 (Proteintech: N-terminal, #17282-1-AP), MNK1 (sc-6965), MNK2 (sc-6964), CDK6 (sc-7961), eIF4G (sc-11373), MCL-1 (#sc-819) (Santa Cruz) or GFP (EVN-AB513, Axxora). All primary antibodies were used at 1: 1,000 dilution. Densitometry analyses were performed using ImageJ software (NIH) and presented as ratio of target band signal intensity to GAPDH band signal intensity. Full immunoblots are presented in Supplementary Fig.10 .
P mnk1 2
Manufactured by Cell Signaling Technology
P-MNK1/2 is an antibody that recognizes the phosphorylated forms of MAP kinase-interacting serine/threonine-protein kinase 1 and 2 (MNK1 and MNK2). MNKs are serine/threonine protein kinases that are activated by phosphorylation through the ERK and p38 MAPK pathways. This antibody can be used to detect the phosphorylation status of these kinases.
Automatically generated - may contain errors
Lab products found in correlation
Phosphatase inhibitor cocktails 2 and 3, by Merck Group (2 mentions)
N myc, by Cell Signaling Technology (2 mentions)
Protease inhibitor, by Merck Group (2 mentions)
Nupage 4 12 bis tris gradient gel, by Thermo Fisher Scientific (2 mentions)
Eif4a, by Cell Signaling Technology (2 mentions)
C myc, by Cell Signaling Technology (2 mentions)
Mcl 1, by Santa Cruz Biotechnology (1 mentions)
Eif4g, by Santa Cruz Biotechnology (1 mentions)
Ab8245, by Abcam (1 mentions)
Sc 9976, by Cell Signaling Technology (1 mentions)
Gapdh, by Abcam (1 mentions)
2 protocols using p mnk1 2
1
Immunoblot analysis of cellular signaling
2
Protein Extraction and Immunoblotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lysed in lysis buffer (25 mM Tris pH 8.0, 125 mM NaCl, 1 mM MgCl2, 1% Triton X-100, 1 mM sodium orthovanadate, 1 mM sodium fluoride, 1X protease inhibitor (Sigma-Aldrich), phosphatase inhibitor cocktails #2 and #3 (Sigma-Aldrich), and 1 mM PMSF). Cell lysates were resolved on a Nupage 4–12% Bis-Tris gradient gel (Life Technologies) and probed using antibodies recognizing phospho- and total eIF4E1 (Cell Signaling #9741 and Santa Cruz sc-9976), phospho- and total ERK (#9101 and #9102, Cell Signaling), c-Myc (#9402), n-Myc (#9405), p-MNK1/2 (#2111), dicer (#3363), eIF4A (#2013) (Cell Signaling), GAPDH (ab8245), BTK (ab54219), YY1 (ab12132) (Abcam), eIF4E3 (Proteintech: N-terminal, #17282-1-AP), MNK1 (sc-6965), MNK2 (sc-6964), CDK6 (sc-7961), eIF4G (sc-11373), MCL-1 (#sc-819) (Santa Cruz) or GFP (EVN-AB513, Axxora). All primary antibodies were used at 1:1,000 dilution. Densitometry analyses were performed using ImageJ software (NIH) and presented as ratio of target band signal intensity to GAPDH band signal intensity. Full immunoblots are presented in Supplementary Fig. 10 .
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