4E-BP1 Antibody - #AF6432

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Product: 4E-BP1 Antibody
Catalog: AF6432
Description: Rabbit polyclonal antibody to 4E-BP1
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 18kDa; 13kD(Calculated).
Uniprot: Q13541
RRID: AB_2835257

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 100ul $280 In stock
 200ul $350 In stock

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(82%)
Clonality:
Polyclonal
Specificity:
4E-BP1 Antibody detects endogenous levels of total 4E-BP1.
RRID:
AB_2835257
Cite Format: Affinity Biosciences Cat# AF6432, RRID:AB_2835257.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

4E-BP1; 4EBP1; 4EBP1_HUMAN; BP 1; eIF4E binding protein 1; eIF4E-binding protein 1; Eif4ebp1; Eukaryotic translation initiation factor 4E-binding protein 1; PHAS-I; PHASI; Phosphorylated heat- and acid-stable protein regulated by insulin 1;

Immunogens

Immunogen:
Uniprot:

Q13541(4EBP1_HUMAN) >>Visit HPA database.

Gene(ID):
EIF4EBP1(1978)
Description:
4E-BP1 binds to eIF4E, preventing its assembly into the EIF4F complex and inhibiting cap-dependent translation. Phosphorylation of 4E-BP1 disrupts this binding, activating cap-dependent translation.
Sequence:
MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPGDYSTTPGGTLFSTTPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPMEASQSHLRNSPEDKRAGGEESQFEMDI

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Zebrafish
100
Rabbit
100
Chicken
82
Xenopus
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q13541 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
S5 Phosphorylation
S6 Phosphorylation
S8 Phosphorylation Q13315 (ATM)
T10 Phosphorylation
S12 Phosphorylation
Y34 Phosphorylation
S35 Phosphorylation
T36 Phosphorylation P51449 (RORC) , O75582 (RPS6KA5) , P31751 (AKT2) , P42345 (MTOR)
T37 Phosphorylation P49841 (GSK3B) , Q16539 (MAPK14) , P31749 (AKT1) , P42345 (MTOR) , P06493 (CDK1) , P28482 (MAPK1) , Q5S007 (LRRK2)
T41 Phosphorylation P42345 (MTOR) , P49674 (CSNK1E)
S44 Phosphorylation P42345 (MTOR)
T45 Phosphorylation O75582 (RPS6KA5) , P28482 (MAPK1) , P42345 (MTOR) , P31751 (AKT2) , P51449 (RORC)
T46 Phosphorylation P06493 (CDK1) , Q5S007 (LRRK2) , Q16539 (MAPK14) , P42345 (MTOR) , P31749 (AKT1) , P28482 (MAPK1) , P49841 (GSK3B)
T50 Phosphorylation P49674 (CSNK1E)
Y54 Phosphorylation
K57 Ubiquitination
C62 S-Nitrosylation
R63 Methylation
S65 Phosphorylation P27361 (MAPK3) , P42345 (MTOR) , P28482 (MAPK1) , O75582 (RPS6KA5) , Q8TAI7 (RHEBL1) , P31749 (AKT1) , Q92630 (DYRK2) , Q9P1W9 (PIM2) , P06493 (CDK1) , Q16539 (MAPK14)
T68 Phosphorylation
K69 Ubiquitination
T70 Phosphorylation P28482 (MAPK1) , P06493 (CDK1) , Q16539 (MAPK14) , P42345 (MTOR) , P41279 (MAP3K8)
T77 Phosphorylation
T82 Phosphorylation
S83 Phosphorylation P42345 (MTOR) , P06493 (CDK1) , P28482 (MAPK1)
S85 Phosphorylation
S86 Phosphorylation
S94 Phosphorylation Q13315 (ATM) , Q13535 (ATR)
S96 Phosphorylation
S101 Phosphorylation Q92630 (DYRK2) , P42345 (MTOR)
S112 Phosphorylation P19784 (CSNK2A2) , P68400 (CSNK2A1) , Q13315 (ATM)

Research Backgrounds

Function:

Repressor of translation initiation that regulates EIF4E activity by preventing its assembly into the eIF4F complex: hypophosphorylated form competes with EIF4G1/EIF4G3 and strongly binds to EIF4E, leading to repress translation. In contrast, hyperphosphorylated form dissociates from EIF4E, allowing interaction between EIF4G1/EIF4G3 and EIF4E, leading to initiation of translation. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways.

PTMs:

Phosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70, corresponding to the hyperphosphorylated form, is regulated by mTORC1 and abolishes binding to EIF4E.

Ubiquitinated: when eIF4E levels are low, hypophosphorylated form is ubiquitinated by the BCR(KLHL25) complex, leading to its degradation and serving as a homeostatic mechanism to maintain translation and prevent eIF4E inhibition when eIF4E levels are low. Not ubiquitinated when hyperphosphorylated (at Thr-37, Thr-46, Ser-65 and Thr-70) or associated with eIF4E.

Subunit Structure:

Hypophosphorylated EIF4EBP1 competes with EIF4G1/EIF4G3 to interact with EIF4E; insulin stimulated MAP-kinase (MAPK1 and MAPK3) or mTORC1 phosphorylation of EIF4EBP1 causes dissociation of the complex allowing EIF4G1/EIF4G3 to bind and consequent initiation of translation. Interacts (via TOS motif) with RPTOR; promoting phosphorylation by mTORC1.

Family&Domains:

The TOS motif mediates interaction with RPTOR, leading to promote phosphorylation by mTORC1 complex.

Belongs to the eIF4E-binding protein family.

Research Fields

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > ErbB signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Genetic Information Processing > Translation > RNA transport.

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

References

1). Patchouli alcohol protects against chronic unpredictable mild stress-induced depressant-like behavior through inhibiting excessive autophagy via activation of mTOR signaling pathway. BIOMEDICINE & PHARMACOTHERAPY, 2020 (PubMed: 32244196) [IF=7.5]

Application: WB    Species: rat    Sample: hippocampus

Fig. 4. |The effect of PA on the level of expression of p-4E-BP-1 protein in the hippocampus. (A) The effect of PA on p-4E-BP-1 and 4E-BP-1 protein levels in hippocampus were investigated by western blot analysis.
2). α-Hydroxyisocaproic Acid Decreases Protein Synthesis but Attenuates TNFα/IFNγ Co-Exposure-Induced Protein Degradation and Myotube Atrophy via Suppression of iNOS and IL-6 in Murine C2C12 Myotube. Nutrients, 2021 (PubMed: 34371902) [IF=5.9]

Application: WB    Species: Mice    Sample:

Figure 2 The effects of HICA on the intracellular signaling pathways. A typical image for a capillary immunoassay is shown (A). The phosphorylation levels of (B) p70S6K and 4E-BP1; (C) AMPK, ACC, and ULK1; (D) ERK1/2; (E) p38MAPK; and (F) eEF2 are shown. The phosphorylation is normalized to the total protein expression. The β-tubulin content in the lysate was measured as a loading control (G). The time course of these experiments is shown in the upper region. DM: differentiation medium, DMEM: Dulbecco’s modified Eagle’s medium, and w/o AA: without amino acids. Data are displayed as the means ± SD, and n = 4 for each group in all bar graphs. * p < 0.05 and ** p < 0.01 vs. the vehicle-treated group.
3). Fluoxetine regulates mTOR signalling in a region-dependent manner in depression-like mice. Scientific Reports, 2015 (PubMed: 26522512) [IF=4.6]

Application: WB    Species: mouse    Sample: mouse

Figure 5. Effect of fluoxetine (20mg/kg) and rapamycin (10mg/kg) on the level of phosphorylated-4E-BP-1 expression in the frontal cortex (A), the hippocampus (B), the amygdala (C) and the hypothalamus (D). The data represented the values of mean± S.E.M. from 5mice/group. # P< 0.05 and ##P< 0.01 vs Control-vehicle group. *P< 0.05 and **P< 0.01 vs CUMS-vehicle group. +P< 0.05 vs CUMS-fluoxetine group. The results of Two-way ANOVA are provided in supplemental materials.
4). mTOR Modulates the Endoplasmic Reticulum Stress-Induced CD4+ T Cell Apoptosis Mediated by ROS in Septic Immunosuppression. MEDIATORS OF INFLAMMATION, 2022 (PubMed: 35915740) [IF=4.6]

Application: WB    Species: Mouse    Sample:

Figure 3 Expression levels of mTOR pathway proteins, ERS-associated proteins, and apoptosis-associated proteins in WT, CLP, and CLP+4-PBA mouse groups. After purifying the CD4+ T cells from mouse spleen lymphocytes, whole cell lysates were assessed for the protein expression of (a) patterns of mTOR pathway proteins, including mTOR, P-mTOR, downstream effectors p70s6k, p-p70s6k, 4EBP, and P-4EBP; (b) ERS-associated proteins, including GRP78 and CHOP; (c) apoptosis-associated proteins, including caspase-3, Bax, and Bcl-2. The protein expression was detected by immunoblotting. Data are mean ± SD. n = 4 biologically independent experiments (one-way ANOVA Tukey's post hoc test). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
5). Rapamycin inhibits B16 melanoma cell viability invitro and invivo by inducing autophagy and inhibiting the mTOR/p70‑S6k pathway. Oncology letters, 2024 (PubMed: 38385108) [IF=2.9]

Application: WB    Species: Mouse    Sample: B16 cells

Figure 5. RAPA inhibits the mTOR/p70-S6k signaling pathway in B16 cells. B16 cells were treated with 0, 1, 10 and 100 nM RAPA for 48 h. (A) Western blotting results showed that RAPA inhibited the expression of p-mTOR and p-p70-S6k compared with in the control group. Statistical analysis of the (B) p-mTOR/mTOR ratio, (C) the p-p70-S6k/p70-S6k ratio and (D) the p-4E-BP1/4E-BP1 ratio at the protein level. Data are presented as the mean ± SD (n=3). *P

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