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Phospho eif4e

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Phospho-eIF4E is a lab equipment product from Cell Signaling Technology. It is a reagent used to detect the phosphorylation of the eukaryotic translation initiation factor 4E (eIF4E) protein, which is involved in the regulation of protein synthesis.

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Lab products found in correlation

Phospho 4ebp1, by Cell Signaling Technology (3 mentions) Phospho akt, by Cell Signaling Technology (2 mentions) Phospho stat3, by Cell Signaling Technology (2 mentions) 4e bp1, by Cell Signaling Technology (2 mentions) Prostaglandin e2 pge2, by Cayman Chemical (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Pharmingen, by BD (1 mentions) Caspase 8, by Cell Signaling Technology (1 mentions) Cobalt 2 chloride cocl2, by Merck Group (1 mentions) Phospho p70s6k1, by Cell Signaling Technology (1 mentions) Phospho cdc25c, by Cell Signaling Technology (1 mentions) Cyclin b1, by Santa Cruz Biotechnology (1 mentions) Phospho chk1, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Mg132, by Merck Group (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Caspase 9, by Cell Signaling Technology (1 mentions) Phospho cdc2, by Cell Signaling Technology (1 mentions) Cyclin a, by Santa Cruz Biotechnology (1 mentions) Phospho mtor, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

EIF4E (87 citations) Phospho-eIF4E(Ser209 (8 citations) Anti-phospho-eIF4E (5 citations) Phospho-eIF4E (Ser209) antibody (2 citations) Phospho-eIF4E (S209) (2 citations)

7 protocols using phospho eif4e

1

BDNF Signaling Pathway Analysis

Cited 1 time
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The BDNF antibodies were purchased from Developmental Studies Hybridoma Bank at the University of Iowa (mouse #9; Iowa City, IA, United States) and Sigma–Aldrich (rabbit; St. Louis, MO, United States). Phospho-eIF4E, GAPDH, and trkB antibodies were obtained from Cell Signaling Technology (Danvers, MA, United States). PAR2 agonist, 2-aminothiazol-4-yl-LIGRL-NH2 (2at-LIGRL), was synthesized as described previously (Boitano et al., 2011 (link)). Human recombinant BDNF was purchased from R&D Systems (Minneapolis, MN, United States). Prostaglandin E2 (PGE2) was purchased from Cayman chemicals (Ann Arbor, MI, United States). All other chemicals were attained from ThermoFisher Scientific (Waltham, MA, United States).
Moy J.K., Khoutorsky A., Asiedu M.N., Dussor G, & Price T.J. (2018). eIF4E Phosphorylation Influences Bdnf mRNA Translation in Mouse Dorsal Root Ganglion Neurons. Frontiers in Cellular Neuroscience, 12, 29.
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2

Salternamide A Biochemical Assay

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Salternamide A (SA, Figure 1A) was dissolved in 100% DMSO and stored at −20 °C for subsequent analysis. Cobalt (II) chloride (CoCl2) and MG132 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies for HIF-1α, Akt, phospho-Akt (Thr308), PI3K, phospho-PI3K (Tyr458/199), RPS6, phospho-RPS6 (Ser235/236), p70S6K1, phospho-p70S6K1 (Thr389), phospho-STAT3 (Tyr705), mTOR, phospho-mTOR (Ser2448), 4E-BP1, phospho-4E-BP1 (Thr37/46), eIF4E, phospho-eIF4E (Ser209), phospho-CDC2 (Thr161), CDC25C, phospho-CDC25C (Ser216), Chk1, phospho-Chk1 (Ser345), phospho-Chk2 (Thr168), caspase-3, caspase-8, caspase-9, cleaved caspase-3, cleaved caspase-8, and LC3B were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies for ERK 1/2, phospho-ERK 1/2 (Thr202/Tyr204), STAT3, CDC2, cyclin B1, cyclin A, Bcl-2, and Bcl-xL were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for VHL, PARP, cleaved PARP, and Bim were purchased from BD Pharmingen™ (BD Biosciences, San Jose, CA, USA). Hsp90 antibody was purchased from Stressgen Bioreagents (Ann Arbor, MI, USA).
Bach D.H., Kim S.H., Hong J.Y., Park H.J., Oh D.C, & Lee S.K. (2015). Salternamide A Suppresses Hypoxia-Induced Accumulation of HIF-1α and Induces Apoptosis in Human Colorectal Cancer Cells. Marine Drugs, 13(11), 6962-6976.
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3

Western Blot Analysis of Signaling Proteins

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Whole cell extracts of cultured cells were prepared in RIPA lysis buffer supplemented with phosphatase and protease inhibitors, and separated on SDS-PAGE gel. The following antibodies and dilution factors were used: phospho-eIF4E (1:1000, Cell Signaling), total eIF4E (1:1000, Santa Cruz Biotechnology), phospho-MNK1 (1:1000, Cell Signaling), total MNK1 (1:1000, Cell Signaling), phospho-ERK1/2 (1:1000, Cell Signaling), total ERK1/2 (1:2000, Santa Cruz), BRD4 (1:1000, Abcam), Rac1 (1:2000, EMD Millipore), RacGAP1 (1:2000, Santa Cruz), and HSP90 (1:3000, Santa Cruz). Blocking agent was 5% bovine serum albumin (BSA). Secondary anti-mouse IgG (A4416) and anti-rabbit IgG (A6667) antibodies were purchased from Sigma and used at a 1:3000 dilution factor. When necessary, membrane was stripped using Restore Western Blot Stripping Buffer (Thermo Fisher Scientific).
Pham T.N., Kumar K., DeCant B.T., Shang M., Munshi S.Z., Matsangou M., Ebine K, & Munshi H.G. (2018). Induction of MNK kinases-dependent eIF4E phosphorylation by inhibitors targeting BET proteins limits efficacy of BET inhibitors. Molecular cancer therapeutics, 18(2), 235-244.
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4

Molecular Mechanisms of LPS-Induced Inflammation

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Most chemical reagents including LPS, CGP57380, pentobarbital sodium, DMSO, paraformaldehyde, and H&E staining solution were purchased from Sigma-Aldrich China Branch (Shanghai, China). The cell culture reagents including DMEM, penicillin, streptomycin, trypsin, and phosphate-buffered saline were purchased from Life Technologies of Thermo Fisher Scientific (China) (Shanghai, China). Fetal bovine serum (FBS) was purchased from EVERY GREEN by Zhejiang Tianhang Biotechnology (Hangzhou, Zhejiang, China). The antibodies against eIF4E, phospho-eIF4E, phospho-ERK, phospho-JNK and phospho-p38 MAPK used in Western blotting were purchased from Cell signaling Technology (Beverly, MA, USA). The FITC-labeled anti-mouse Ly-6G (Gr-1) antibody was obtained from eBioscience (Grand Island, NY). The GAPDH antibody was purchased from Proteintech (Wuhan, Hubei, China). The secondary antibody (horseradish peroxidase-conjugated goat anti-rabbit IgG antibody) was obtained from Mei5 Biotechnology (Beijing, China). The enhanced chemiluminescence (ECL) kit was purchased from Thermo Fisher Scientific (China) (Shanghai, China).
Gao J., Teng L., Yang S., Huang S., Li L., Zhou L., Liu G, & Tang H. (2021). MNK as a potential pharmacological target for suppressing LPS-induced acute lung injury in mice. Biochemical Pharmacology, 186, 114499.
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5

Immunoblotting Procedures and Antibodies

Cited 2 times
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Immunoblotting was conducted as previously described (15 (link),25 (link)). The primary antibodies used for western blot were c-Myc (SC-40, 1:500, Santa Cruz Technology); phospho-c-Myc (#13748, c-Myc-pSer62, 1:500; Cell Signaling Technology); phospho-c-Myc (Y011034, c-Myc-pThr58, 1:1000; Applied Biological Materials Inc.); RPA2 (Clone NA18, 1:100, Calbiochem/EMD Millipore); anti-β-actin (Clone AC-74, 1:50000, Sigma-Aldrich); anti-CHK1 (G-4, 1:200, Santa Cruz Technology); phospho-CHK1 antibody (#133D3, CHK1-pSer345, 1:500, Cell Signaling Technology); CDC45 (G-12 sc55569, 1:200, Santa Cruz Technology); γ-H2Ax (ser139, clone JBC301, 1:500, Millipore); rabbit polyclonal antibody phosphor RPA32 Ser4/Ser8 (BL647, 1: 1000, Bethyl; Histone H3 (#9715, 1:1000, Cell Signaling Technology); PPP2R5A (ab89621,1:1000, Abcam); eIF4E (A2162, 1:1000, Abclonal); Phospho-eIF4E (#9741, eIF4E-Ser209, 1:1000, Cell Signaling Technology); 4E-BP1 (#9452, 1:1000, Cell Signaling Technology); Phospho-4E-BP1 (#2855, 4E-BP1-Thr36/47, 1:1000, Cell Signaling Technology); Phospho-p70S6K antibody (#9205, p70S6K-Thr389, 1:1000, Cell Signaling Technology); p70S6K (#9207, 1:1000, Cell Signaling Technology); Cdc45 (H-300 clone, SC20685, 1:50, Santa Cruz Technology); and phospho-Histone H3 (Ser10); (#9706, 1:100, Cell Signaling Technology).
Qiu Z., Fa P., Liu T., Prasad C.B., Ma S., Hong Z., Chan E.R., Wang H., Li Z., He K., Wang Q.E., Williams T.M., Yan C., Sizemore S.T., Narla G, & Zhang J. (2020). A genome-wide pooled shRNA screen identifies PPP2R2A as a predictive biomarker for the response to ATR and CHK1 inhibitors. Cancer research, 80(16), 3305-3318.
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6

Apoptosis Pathway Antibody Protocol

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Antibodies against cleaved caspase-3, 8, 9, PARP, Bax, Bak, Bad, Bim, Puma, Bcl-2, Bcl-xL, Mcl-1, DR5, DR4 and phospho-eIF4E were from Cell Signaling (Danvers, MA, USA) and Cox4 antibody was from Abcam (Cambridge, UK); cytochrome C antibody was from BD Biosciences (San Diego, CA, USA); MG-132 and antibodies for Sp1, α-tubulin and actin were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA); DAPI, CHX and Mith was from Sigma-Aldrich Chemical Co. (St Louis, MO, USA); zVAD-fmk, a pancaspase inhibitor, was from R&D Systems (Minneapolis, MN, USA).
Choi E.S., Nam J.S., Jung J.Y., Cho N.P, & Cho S.D. (2014). Modulation of specificity protein 1 by mithramycin A as a novel therapeutic strategy for cervical cancer. Scientific Reports, 4, 7162.
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7

Western Blot Analysis of Signaling Pathways

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Whole protein extraction and Western blot analysis were done as previously described.[18 (link)] Membranes were probed with the following primary antibodies from Cell Signaling Technology (Danvers): phospho-Akt (Ser473), phospho-ribosomal protein S6 (RPS6; Ser235/236), phospho-4EBP1 (Thr37/46), phospho-eIF4E (Ser209), phospho-STAT3 (Tyr705, clone D3A7), phospho-STAT6 (Tyr641), Akt, RPS6, 4EBP1, eIF4E, STAT3 (clone 79D7) and STAT6 (clone D3H4). Horseradish peroxidase-labeled anti-mouse IgG (Sigma) and anti-rabbit IgG (Cell Signaling Technology) were used as secondary antibodies. Ratio between phosphorylated and total protein levels was calculated and relative protein quantification in treated versus control extracts was conducted with Image Gauge software (Fujifilm). Equal protein loading was confirmed by re-probing membranes with anti-β-actin or anti-α-tubulin antibodies (Sigma).
Rosich L., Montraveta A., Xargay-Torrent S., López-Guerra M., Roldán J., Aymerich M., Salaverria I., Beà S., Campo E., Pérez-Galán P., Roué G, & Colomer D. (2014). Dual PI3K/mTOR inhibition is required to effectively impair microenvironment survival signals in mantle cell lymphoma. Oncotarget, 5(16), 6788-6800.
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