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Eif4b

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EIF4B is a protein that plays a role in the initiation of protein synthesis. It is involved in the regulation of translation, a process that converts mRNA into proteins. EIF4B functions as part of the eukaryotic translation initiation complex.

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Lab products found in correlation

Anti puromycin, by Merck Group (2 mentions) P eif4b, by Cell Signaling Technology (2 mentions) Eif4g, by Cell Signaling Technology (2 mentions) Eif4e, by Cell Signaling Technology (2 mentions) P eif4e, by Cell Signaling Technology (2 mentions) P eif4g, by Cell Signaling Technology (2 mentions) 4e bp1, by Cell Signaling Technology (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions) Phosphorylated p70s6k 389, by Cell Signaling Technology (1 mentions) Phosphorylated s6 240 244, by Cell Signaling Technology (1 mentions) Peroxidase labeled anti mouse igg2a, by Jackson ImmunoResearch (1 mentions) Doxycycline, by Selleck Chemicals (1 mentions) Erk1 2, by ABclonal (1 mentions) Ezview red anti flag m2 affinity gel, by Merck Group (1 mentions) Sumo1, by ABclonal (1 mentions) Glutathione sepharose 4b, by GE Healthcare (1 mentions) Sodium butyrate, by Merck Group (1 mentions) Clonexpress 2 one step cloning kit, by Vazyme (1 mentions) Hiscript 2 q rt supermix for qpcr gdna wiper, by Vazyme (1 mentions) Phosphorylated s6, by Cell Signaling Technology (1 mentions)

6 protocols using eif4b

1

Phosphorylation Signaling Pathway Analysis

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Antibodies targeting 4E-BP1 (1:1000, #9644), eEF2 (1:3000, #2332), phosphorylated eEF2(56) (1:1000, #2331), eIF4B (1:1000, #3592), phosphorylated eIF4B (422) (1:1000, #3591), MKK4 (1:1000, #3346), phosphorylated MKK4 (257) (1:1000, #4514), p38 (1:1000, #9212), phosphorylated p38 (180/182) (1:1000, #9211), p70S6K (1:1000, #9234), phosphorylated p70S6K (389) (1:1000, #2708), S6 (1:1000, #2217), and phosphorylated S6 (240/244) (1:1000, #5364) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-puromycin (1:5000, 12D10) was purchased from MilliporeSigma (Burlington, MA, USA). Peroxidase-labeled anti-mouse IgG2a (1:20,000, 115-035-206) was obtained from Jackson ImmunoResearch Laboratories, Inc (West Grove, PA, USA). Peroxidase-labeled anti-rabbit (1:5000, PI-1000) was obtained from Vector Labs Inc. (Burlingame, CA, USA).
Zhu W.G., Hibbert J.E., Lin K.H., Steinert N.D., Lemens J.L., Jorgenson K.W., Newman S.M., Lamming D.W, & Hornberger T.A. (2021). Weight Pulling: A Novel Mouse Model of Human Progressive Resistance Exercise. Cells, 10(9), 2459.
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2

Comprehensive Antibody Validation Protocol

Cited 1 time
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Anti-RxRxxS*/T*(#10001S), RSK1 (#9333S), RSK2 (#5528S), p-eIF4B (#3591S), eIF4B (#3592S) antibodies were ordered from Cell Signaling Technology. HRP anti-HA (#901519), HRP anti-Flag (#637311) antibodies were purchased from BioLegend. pRSK1 (Thr359/Ser363) (#AP0539), pRSK1 (Ser380) (#AP1147), RSK3 (#A16305), ERK1/2 (#A10613), and SUMO1 (#A19121) were purchased from ABclonal. Anti-RTA (ORF50) monoclonal mouse antibody was given by Dr. Ke Lan (Wuhan University, China) [34 (link)]. Anti-ORF65 monoclonal mouse antibody was given by Dr. Shou-Jiang Gao (University of Pittsburgh, USA) [35 (link)]. Monoclonal antibodies against ORF45 and ORF52 and polyclonal antibodies against K3, K8, and ORF55 were described previously [36 (link)–38 (link)]. EZview Red ANTI-FLAG M2 Affinity Gel (#F2426) and sodium butyrate (#B5887) were ordered from Sigma, Doxycycline (#S4163) was ordered from Selleck, Glutathione Sepharose 4B (#17-0756-01) were ordered from GE Healthcare. ClonExpress II One Step Cloning Kit (#C122-01) and HiScript II Q RT SuperMix for qPCR (+gDNA wiper) (#R223-01) were purchased from Vazyme Biotech. Lipofectamine 3000 (#3000015) were purchased from Thermo Fisher Scientific.
Liu Z., Liu C., Wang X., Li W., Zhou J., Dong P., Xiao M.Z., Wang C., Zhang Y., Fu J., Zhu F, & Liang Q. (2021). RSK1 SUMOylation is required for KSHV lytic replication. PLoS Pathogens, 17(12), e1010123.
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3

Western Blot Analysis of Protein Signaling

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Cell lysates were run on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was then immunoblotted with specific primary antibodies: eIF4E, eIF4B, S6, phosphorylated eIF4E, phosphorylated eIF4B (S406 and S422), phosphorylated S6, (Cell Signaling Technology, Danvers, MA), and β-actin (Sigma). Antibodies were visualized using the ECL chemilu-minescent detection system (Fisher Scientific) using a UVP/Analytik-Jena Imager (Analytik Jena, Upland, CA). Density of each protein band was determined by densitometric analysis using the VisionWorks image analysis software (Analytik Jena). Differences in protein expression are determined compared to the control. β-actin levels were determined for each treatment to monitor protein loading. The density of each protein band was normalized to β-actin to correct for minor differences in protein loading among the lanes.
Ruff A., Lewis M, & Whalen M. (2022). Organotin and organochlorine toxicants activate key translational regulatory proteins in human immune cells. Archives of toxicology, 97(2), 469-493.
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4

Western Blotting Protein Analysis

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Western blotting analyses were performed as described previously (Marabita et al., 2016 (link)). Antibodies for P-Akt, Akt, eIF4E, P-eIF4E, P-eIF4B, eIF4B, eIF4A, eIF4H, eIF4G, P-eIF4G, 4E-BP1, P-S6, S6 were taken from Cell Signaling. Actin was from Santa Cruz, and puromycin from Millipore. All quantifications of the western blots were done on at least four different blots for each protein, in each condition. Differences between groups were assessed using Student's t-test. Significance was defined as a value of P < 0.05 (95% confidence).
Pereira M.G., Dyar K.A., Nogara L., Solagna F., Marabita M., Baraldo M., Chemello F., Germinario E., Romanello V., Nolte H, & Blaauw B. (2017). Comparative Analysis of Muscle Hypertrophy Models Reveals Divergent Gene Transcription Profiles and Points to Translational Regulation of Muscle Growth through Increased mTOR Signaling. Frontiers in Physiology, 8, 968.
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5

Protein Expression Analysis Protocol

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Primary antibodies from Cell Signaling Technology were used to analyze the expression of important proteins, Bcl-2 (catalog #4223S), Bax (catalog #2772S), Bcl-xL (catalog #2764S), eIF4e (catalog #9742S), peIF4e (catalog #9741S), 4EBP1 (catalog #9452S), p4EBP1 (catalog #2855S), mTOR (catalog #4517S), p-mTOR (catalog #5536S), eIF4B (catalog #3592S), peIF4B (catalog #3591S), eIF4G (catalog #2498S), peIF4G (catalog #2441S), p18 (catalog # 2896S) and p21 (catalog #2947S). Cleaved PARP (sc-8007) and Cyclin D1 (sc-753) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). β-actin antibody was obtained from Sigma (catalog #A2228; St. Louis, MO, USA). The horseradish peroxidase conjugated rabbit (catalog #4011) and mouse (catalog #4021) secondary antibodies were procured from Promega (Madison, WI, USA).
Kumari S., Sikander M., Malik S., Tripathi M.K., Hafeez B.B., Yallapu M.M., Chauhan S.C., Khan S, & Jaggi M. (2021). Steviol Represses Glucose Metabolism and Translation Initiation in Pancreatic Cancer Cells. Biomedicines, 9(12), 1814.
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6

Western Blot Antibody Validation

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Antibodies targeting 4E-BP1(1:2000, #9644), phospho 4E-BP1(37/46) (1:1000, #2855), eEF2 (1:3000, #2332), phospho eEF2(57) (1:1000, #2331), eIF2α (1:1000, #5324), phospho eIF2α(51) (1:1000, #3398), eIF4B (1:1000, #3592), phospho eIF4B(422) (1:1000, #3591), RPS6 (1:1000, #2217) and phospho RPS6(235/236) (1:1000, #2211) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-puromycin (1:5000, 12D10) was purchased from MilliporeSigma. Anti-complement C3 (1:3000, 855463) was purchased from MP Biomedicals (Santa Ana, CA, USA). Peroxidase-labelled anti-mouse IgG2a (1:50 000, 115-035-206) was from Jackson ImmunoResearch Laboratories, Inc (West Grove, PA, USA). Peroxidase-labelled anti-rabbit (1:5000, PI-1000), peroxidase-labelled anti-mouse (1:5000, PI-2000) and peroxidase-labelled anti-goat (1:5000, PI-9500) were obtained from Vector Labs Inc. (Burlingame, CA, USA).
Lin K.H., Wilson G.M., Blanco R., Steinert N.D., Zhu W.G., Coon J.J, & Hornberger T.A. (2021). A deep analysis of the proteomic and phosphoproteomic alterations that occur in skeletal muscle after the onset of immobilization. The Journal of physiology, 599(11), 2887-2906.
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