Product: Phospho-4E-BP1 (Thr37/Thr46) Antibody
Catalog: AF3830
Description: Rabbit polyclonal antibody to Phospho-4E-BP1 (Thr37/Thr46)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 17kD; 13kD(Calculated).
Uniprot: Q13541
RRID: AB_2847144

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 100ul $350 In stock
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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
Clonality:
Polyclonal
Specificity:
Phospho-4E-BP1 (Thr37/Thr46) Antibody detects endogenous levels of 4E-BP1 only when phosphorylated at Thr37/46.
RRID:
AB_2847144
Cite Format: Affinity Biosciences Cat# AF3830, RRID:AB_2847144.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
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:

A synthesized peptide derived from human 4E-BP1 around the phosphorylation site of Thr37/46.

Uniprot:
Gene(ID):
Sequence:
MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPGDYSTTPGGTLFSTTPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPMEASQSHLRNSPEDKRAGGEESQFEMDI

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). Kaempferol Improves Exercise Performance by Regulating Glucose Uptake, Mitochondrial Biogenesis, and Protein Synthesis via PI3K/AKT and MAPK Signaling Pathways. Foods (Basel, Switzerland), 2024 (PubMed: 38611372) [IF=5.2]

Application: IF/ICC    Species: Mouse    Sample:

Figure 6 Protein expression/phosphorylation of downstream maker targeted to biological functions in C2C12 myotubes. Protein expression of mtTFA (A) and GLUT4 (B) and the phosphorylation of 4EBP1 (C) and p70S6K (D) were detected with a high-content imaging analysis system. Left panel: representative images, bar size: 50 μm. The assays were performed in triplicate with three technical replicates.

2). Protective effects and potential mechanisms of (2-Carboxyethyl) dimethylsulfonium Bromide (Br-DMPT) on gill health status of on-growing grass carp (Ctenopharyngodon idella) after infection with Flavobacterium columnare. FISH & SHELLFISH IMMUNOLOGY, 2020 (PubMed: 32771611) [IF=4.7]

Application: WB    Species: grass carp    Sample: gill

Fig. 8. |Western blot analysis of TOR protein phosphorylation (p-TOR Ser2448) (A), S6K1 protein phosphorylation (p-S6K1 Thr-389) (B), 4EBP1 protein phosphorylation (Thr-37/46) (C) in the gill of on-growing grass carp fed diets containing graded levels of Br-DMPT for 60 days, then infected with F. columnare for 3 days.Data represent means of six fish in each group, error bars indicate S.D. Values having different letters are significantly different (P < 0.05).

3). 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

4). L-Leucine stimulates proliferation and casein secretion in porcine mammary epithelial cells through mTOR-p70S6K-4EBP1 signalling pathway. , 2022

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