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P-MNK1 is a protein phosphorylation detection antibody that specifically recognizes the phosphorylated form of MKNK1 (MAP Kinase Interacting Serine/Threonine Kinase 1). This antibody can be used to detect the activation state of MKNK1 in samples.

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Lab products found in correlation

Erk1 2, by Cell Signaling Technology (2 mentions) P erk1 2, by Cell Signaling Technology (2 mentions) Mini protean tgx stain free gel, by Bio-Rad (2 mentions) Spark plate reader, by Tecan (2 mentions) Eif4g, by Cell Signaling Technology (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Chemidoc mp imaging system, by Bio-Rad (2 mentions) Eif4e, by Abcam (2 mentions) P eif4e, by Abcam (2 mentions) P eif4g, by Cell Signaling Technology (2 mentions) Alphalisa surefire ultra p eif4e ser209 assay kits, by PerkinElmer (2 mentions) Ecl western blotting detection reagent, by GE Healthcare (1 mentions) P atf2, by Cell Signaling Technology (1 mentions) Eif4e, by Cell Signaling Technology (1 mentions) P eif4e, by Cell Signaling Technology (1 mentions) P p90rsk, by Cell Signaling Technology (1 mentions) Ab17942, by Abcam (1 mentions) Hsp27, by Cell Signaling Technology (1 mentions) Phosstop and complete phosphatase protease inhibitor cocktails, by Roche (1 mentions) P hsp27 s82, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

P-MNK1/2 (2 citations)

5 protocols using p mnk1

1

Western Blot Analysis of Cell Signaling Pathways

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Total protein
extracts were generated using lysis buffer (50 mM Tris-HCl, pH 7.4,
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1
mM EDTA) supplemented with PhosSTOP and Complete Phosphatase/Protease
Inhibitor Cocktails (Roche Diagnostics GmbH, Mannheim, Germany). Protein
extracts (20–25 μg per sample) were loaded onto sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels
and transferred electrophoretically to poly(vinylidene fluoride) (PVDF)
membranes. The following primary antibodies were used at a dilution
of 1:1000: ERK1/2 (p44/42) (ab17942, Abcam), p-ERK1/2 (p-p44/42) (Thr202/Tyr204)
(9101, Cell Signaling), MNK1 (2195, Cell Signaling), p-MNK1 (Thr197/202)
(2111, Cell Signaling), eIF4E (9742, Cell Signaling), p-eIF4E (p-Ser
209) (9741, Cell Signaling or NBP2-66802, Novus Biologicals), p38alpha
(9218, Cell Signaling), p38beta (2339, Cell Signaling), p-p38 (4511,
Cell Signaling), p-ATF2 (27934, Cell Signaling), Hsp27 (2402, Cell
Signaling), p-Hsp27 (S82) (2401, Cell Signaling), and p-p90 RSK (Thr573)
(9346, Cell Signaling). The primary HRP-conjugated antibody anti-β-actin
(Calbiochem) was used at a dilution of 1:20 000. Anti-mouse
and anti-rabbit HRP secondary antibodies were from Pierce and used
at a dilution of 1:10 000. Immunodetection of proteins was
performed using ECL Western Blotting Detection Reagents (GE Healthcare,
Buckinghamshire, U.K.).
Bou-Petit E., Hümmer S., Alarcon H., Slobodnyuk K., Cano-Galietero M., Fuentes P., Guijarro P.J., Muñoz M.J., Suarez-Cabrera L., Santamaria A., Estrada-Tejedor R., Borrell J.I, & Ramón y Cajal S. (2022). Overcoming Paradoxical Kinase Priming by a Novel MNK1 Inhibitor. Journal of Medicinal Chemistry, 65(8), 6070-6087.
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2

Quantification of eIF4E Phosphorylation in Neurons

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Cortical neurons and brain tissue were homogenized in lysis buffer containing NaCl 137 mM, KCl, 2.7 mM, Na2HPO4 10 mM, KH2PO4, 1.8 mM, EDTA 5 mM, Triton 1%, and complete protease and phosphatase inhibitors (Roche Applied Science). Immunoblotting was done with HRP-conjugated secondary antibodies and Pierce ECL Western Blotting Substrate. The following primary antibody was used in this study: p-eIF4E (Abcam, ab76256 1:1000), eIF4E (Abcam, ab47482 1:1000), p-ERK1/2 (Cell Signaling, 4370S 1:1000), ERK1/2 (Cell Signaling, 4695S 1:1000), p-eIF4G (Cell Signaling, 2441S 1:1000), eIF4G (Cell Signaling, 2498 1:1000), p-MNK1 (Cell Signaling, 2111, 1:1000), MNK1 (Cell Signaling, 2195S, 1:1000), GAPDH (Cell Signaling, 5174 1:2000) and calnexin (Stressgen, SPA-865 1:2000). Loading controls were run on the same gel, and for some experiments Mini PROTEAN® TGX Stain-Free™ Gels (Bio-Rad) were used as loading controls. Signals were acquired using an image analyzer (Bio-Rad, ChemiDoc MP Imaging System) and images were analyzed and prepared using ImageJ.
For additional measurements of eIF4E phosphorylation state, the AlphaLISA SureFire Ultra p-eIF4E (Ser209) Assay Kits (PerkinElmer) were used according to the manufactures protocol. AlphaLISA signals were measured using a Tecan SPARK plate reader on recommended settings.
Hörnberg H., Perez-Garci E., Schreiner D., Hattstatt-Burklé L., Magara F., Baudouin S., Matter A., Nacro K., Pecho-Vrieseling E, & Scheiffele P. (2020). Rescue of oxytocin response and social behavior in a rodent model of autism. Nature, 584(7820), 252-256.
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3

Comprehensive Molecular Profiling of Cellular Signaling

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General tissue culture materials were obtained from VWR International. Antibodies against eIF4E, tubulin, HSP90, ZEB1 and Dicer were obtained from Santa Cruz, while antibodies against p-eIF4E, MNK1, p-MNK1 and Drosha were purchased from Cell Signaling. Anti-GAPDH antibody was from Millipore, anti-E-cadherin antibody was obtained from BD Bioscience, while anti-vimentin antibody was from Abcam. Secondary antibodies were purchased from Sigma. CGP57380 was obtained from Santa Cruz. siRNAs against MNK1 and MNK2 were purchased from Dharmacon, ZEB1 siRNA was obtained from Life Technologies, while eIF4E and hnRNPA1 siRNAs were from Santa Cruz. Aldefluor assay kit was purchased from Stemcell Technologies.
Kumar K., Chow C.R., Ebine K., Arslan A.D., Kwok B., Bentrem D.J., Eckerdt F.D., Platanias L.C, & Munshi H.G. (2015). Differential Regulation of ZEB1 and EMT by MAPK-interacting protein kinases (MNKs) and eIF4E in Pancreatic Cancer. Molecular cancer research : MCR, 14(2), 216-227.
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4

Quantification of eIF4E Phosphorylation in Neurons

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Cortical neurons and brain tissue were homogenized in lysis buffer containing NaCl 137 mM, KCl, 2.7 mM, Na2HPO4 10 mM, KH2PO4, 1.8 mM, EDTA 5 mM, Triton 1%, and complete protease and phosphatase inhibitors (Roche Applied Science). Immunoblotting was done with HRP-conjugated secondary antibodies and Pierce ECL Western Blotting Substrate. The following primary antibody was used in this study: p-eIF4E (Abcam, ab76256 1:1000), eIF4E (Abcam, ab47482 1:1000), p-ERK1/2 (Cell Signaling, 4370S 1:1000), ERK1/2 (Cell Signaling, 4695S 1:1000), p-eIF4G (Cell Signaling, 2441S 1:1000), eIF4G (Cell Signaling, 2498 1:1000), p-MNK1 (Cell Signaling, 2111, 1:1000), MNK1 (Cell Signaling, 2195S, 1:1000), GAPDH (Cell Signaling, 5174 1:2000) and calnexin (Stressgen, SPA-865 1:2000). Loading controls were run on the same gel, and for some experiments Mini PROTEAN® TGX Stain-Free™ Gels (Bio-Rad) were used as loading controls. Signals were acquired using an image analyzer (Bio-Rad, ChemiDoc MP Imaging System) and images were analyzed and prepared using ImageJ.
For additional measurements of eIF4E phosphorylation state, the AlphaLISA SureFire Ultra p-eIF4E (Ser209) Assay Kits (PerkinElmer) were used according to the manufactures protocol. AlphaLISA signals were measured using a Tecan SPARK plate reader on recommended settings.
Hörnberg H., Perez-Garci E., Schreiner D., Hattstatt-Burklé L., Magara F., Baudouin S., Matter A., Nacro K., Pecho-Vrieseling E, & Scheiffele P. (2020). Rescue of oxytocin response and social behavior in a rodent model of autism. Nature, 584(7820), 252-256.
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5

Western Blot Analysis of Protein Signaling

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Protein extracts were prepared and separated on 4-20% Tris-glycine gels and electroblotted onto PVDF membranes as previously described [11 (link), 12 (link), 50 (link)]. Protein levels were assessed using antibodies against p53 (Millipore, Bellerica, MA #05-224), p-ATM (Cell Signaling, Danvers, MA #5883P), p-Chk2 (Cell Signaling, Danvers, MA #2661P), p-Chk1 (Cell Signaling, Danvers, MA #2348P), p38 (Cell Signaling, Danvers, MA #8690), histone γ-H2AX (Cell Signaling, Danvers, MA #7631), p-histone γH2AX (Cell Signaling, Danvers, MA #9718P), p-BRCA1 (Ser1524) (Cell Signaling, Danvers, MA #9009P), p-P38 MAPK (Cell Signaling, Danvers, MA #9216S), mTOR (Cell Signaling, Danvers, MA #2983), p-ATR (Cell Signaling, Danvers, MA #2853P), p-p53 (Cell Signaling, Danvers, MA #9286P), p-MNK1 (cell signaling, #2111S), p-4E-BP1 (Cell Signaling, Danvers, MA #2855S), MDM2 (Santa Cruz Biotechnology, #sc-965), β-actin (Cell Signaling, Danvers, MA #4967). Densitometry was performed using ImageJ analysis software (NIH, Bethesda, MD) as previously described [11 (link), 12 (link), 50 (link)].
Waye S., Naeem A., Choudhry M.U., Parasido E., Tricoli L., Sivakumar A., Mikhaiel J.P., Yenugonda V., Rodriguez O.C., Karam S.D., Rood B.R., Avantaggiati M.L, & Albanese C. (2015). The p53 tumor suppressor protein protects against chemotherapeutic stress and apoptosis in human medulloblastoma cells. Aging (Albany NY), 7(10), 854-867.
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