Phospho-4E-BP1 (Thr45) Antibody - #AF3432

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Product: Phospho-4E-BP1 (Thr45) Antibody
Catalog: AF3432
Description: Rabbit polyclonal antibody to Phospho-4E-BP1 (Thr45)
Application: WB IHC IF/ICC
Cited expt.: WB
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 18kDa; 13kD(Calculated).
Uniprot: Q13541
RRID: AB_2834874

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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:
Phospho-4E-BP1 (Thr45) Antibody detects endogenous levels of 4E-BP1 only when phosphorylated at Threonine 45.
RRID:
AB_2834874
Cite Format: Affinity Biosciences Cat# AF3432, RRID:AB_2834874.
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 Thr45.

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

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.

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). Functional characterization of DLK1/MEG3 locus on chromosome 14q32.2 reveals the differentiation of pituitary neuroendocrine tumors. Aging-US, 2021 (PubMed: 33472171) [IF=3.9]

Application: WB    Species: Rat    Sample: GH3 cell

Figure 5 Effect of anit-DLK1 antibody on the bioactivity of PitNET cell lines. (A) Western blot assay measured the levels of DLK1 and PIT1 in GH3 cell line, MMQ cell line and ATT20 cell line. (B) Anti-DLK1 antibody inhibited the cell viability of GH3 cells in the dose- and time-dependent manner, not MMQ cells or ATT20 cells. (C) Anti-DLK1 antibody inhibited the secretion of GH/IGF-1 in GH3 cells, not PRL in MMQ cells and ACTH in ATT20 cells. (D) Clone forming experiment showed the anti-DLK1 antibody promoted the cell proliferation in GH3 cell line. (E) Confocal experiment showed DLK1 regulated the level of PIT1 in GH3 cell line. (F) Western blot experiment showed Anti-DLK1 antibody activated the mTOR pathway in GH3 cell line. *compare to control group P<0.05 **P<0.01 ***P<0.001.
2). Fluoxetine regulates mTOR signalling in a region-dependent manner in depression-like mice. Scientific Reports, 2015 (PubMed: 26522512) [IF=3.8]

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.
3). Melatonin alleviates endoplasmic reticulum stress and follicular granulosa cell apoptosis by regulating ATF4 to activate mTOR signaling pathway in chickens. Poultry science, 2024 (PubMed: 38583308) [IF=3.8]
4). The DLK1/MEG3 Locus is Related to the Differentiation of Pituitary Neuroendocrine Tumors. Research Square, 2020

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