Product Name: 4E-BP1 pT45
Product Number: AB-PN114
Size: 25 µg      Price:89.00
      $US
Target Full Name: Eukaryotic translation initiation factor 4E-binding protein 1 Thr-45 phosphosite

Target Alias: 4EBP1; EIF4EBP1; Eukaryotic translation initiation factor 4E binding protein 1; Insulin-stimulated EIF-4E binding protein; P/OKCL.6; PHAS1; PHAS-1; PHAS-I; Phosphorylated heat- and acid-stable protein regulated by insulin 1

Product Type Specific: Phosphosite-specific polyclonal antibody for human 4E-BP1 T45 phosphosite

Antibody Code: PN114

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: T45

Protein UniProt: Q13541

Protein SigNET: Q13541

Antibody Type: Polyclonal
Antibody Host Species: Rabbit

Antibody Ig Isotype Clone: IgG

Antibody Immunogen Source: Peptide with amino acid sequence surrounding human 4E-BP1 Thr-45 phosphorylation site (Ser-Thr-pThr-Pro-Gly)

Antibody Immunogen Sequence: HSTpTPG

Antibody Immunogen Description: 6 amino acid phosphopeptide corresponding to the human 4E-BP1 T45 phosphosite

Production Method: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using the epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatogramphy using non-phosphopeptide corresponding to the phosphorylation site.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 0.02% sodium azide and 50% glycerol

Storage Conditions: -20°C

Storage Stability: > 1 year

Product Use: Western blotting, Immunohistochemistry (IHC)

Antibody Dilution Recommended: 2 µg/mL

Antibody Potency: Very strong

Antibody Species Reactivity: Human, Mouse, Rat

Antibody Positive Control: T45 phosphorylated 4E-BP1

Antibody Specificity: Very high

Antibody Cross Reactivity: This antibody may cross-react with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites. Ocassionally, strong cross-reactivities with ~90 kDa and ~160 kDa proteins can be detected.

Related Product 1: 4E-BP1-pT37+pT46 phosphosite-specific antibody (Cat. No.: AB-PN550)

Scientific Background: The protein synthesis inhibitor 4E-BP1 (PHAS1) normally competes with EIF4G1/EIF4G3 to interact with EIF4E to prevent its assembly into the EIF4F complex and thereby inhibits cap-dependent translation. Phosphorylation of 4E-BP1 disrupts this binding, and permits initiation of cap-dependent translation. 4E-BP1 phosphorylation is targeted for regulation by hormones, growth factors and other stimuli that signal through the PI3 kinase/mTORC1 and MAPK pathways. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1 and Ser-65 by ERK1/ERK2. It is also phosphorylated upon DNA damage, probably by ATM or ATR.
Figure 1. Immunoblotting of various cell lines with AB-PN114 antibody at a 500 X dilution. The target protein 4E-BP1 is indicated. Each lane was loaded with 15 µg of cell lysate protein. The max signal count was 13055.


Figure 2. Immunoblotting of various cell lines with AB-PN114 antibodies at a 500 X dilution. The target protein 4E-BP1 is indicated. Each lane was loaded with 15 µg of cell lysate protein. The max signal count was 17132.


References

[1] Dephoure N, Zhou C, Villén J, Beausoleil SA, Bakalarski CE, Elledge SJ, Gygi SP. A quantitative atlas of mitotic phosphorylation. Proc Natl Acad Sci U S A. 2008 Aug 5;105(31):10762-7. PMID: 18669648

[2] Gingras AC, Kennedy SG, O'Leary MA, Sonenberg N, Hay N. 4E-BP1, a repressor of mRNA translation, is phosphorylated and inactivated by the Akt(PKB) signaling pathway. Genes Dev. 1998 Feb 15;12(4):502-13. PMID: 9472019

[3] Brugarolas J, Lei K, Hurley RL, Manning BD, Reiling JH, Hafen E, Witters LA, Ellisen LW, Kaelin WG Jr. Regulation of mTOR function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor suppressor complex. Genes Dev. 2004 Dec 1;18(23):2893-2904. PMID: 15545625

[4] Kumar V, Pandey P, Sabatini D, Kumar M, Majumder PK, Bharti A, Carmichael G, Kufe D, Kharbanda S. Functional interaction between RAFT1/FRAP/mTOR and protein kinase cdelta in the regulation of cap-dependent initiation of translation. EMBO J. 2000 Mar 1;19(5):1087-97. PMID: 10698949.

[5] Moody CA, Scott RS, Amirghahari N, Nathan CO, Young LS, Dawson CW, Sixbey JW. Modulation of the cell growth regulator mTOR by Epstein-Barr virus-encoded LMP2A. J Virol. 2005 May;79(9):5499-506. PMID: 15827164.

[6] Burnett PE, Barrow RK, Cohen NA, Snyder SH, Sabatini DM. RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1432-7. PMID: 9465032.