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Phos 4ebp1

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Phos-4EBP1 is a phosphosite-specific antibody that detects phosphorylation of Threonine 37 and Threonine 46 on the 4E-binding protein 1 (4EBP1) protein. 4EBP1 is a key regulator of protein synthesis and is phosphorylated by mTOR.

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Lab products found in correlation

Phos s6 ribosomal protein, by Cell Signaling Technology (3 mentions) S6 ribosomal protein, by Cell Signaling Technology (3 mentions) Lamin a c, by Cell Signaling Technology (3 mentions) 4e bp1, by Cell Signaling Technology (3 mentions) Gapdh, by Cell Signaling Technology (3 mentions) Phos erk, by Cell Signaling Technology (2 mentions) β actin, by Merck Group (2 mentions) Pgc1α, by Abnova (2 mentions) Cyp2e1, by Abcam (1 mentions) Aldh2, by Proteintech (1 mentions) Anti myeloperoxidase mpo, by Biocare Medical (1 mentions) Hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Phosmapk, by Cell Signaling Technology (1 mentions) Rhodamine conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Anti flag m2 gel, by Merck Group (1 mentions) Ha tag, by Proteintech (1 mentions) Azd8055, by MedChemExpress (1 mentions) His tag, by Proteintech (1 mentions) Gapdh, by Proteintech (1 mentions) Spautin 1, by Merck Group (1 mentions)

5 protocols using phos 4ebp1

1

Western Blot Antibody Reagents for Protein Analysis

Cited 1 time
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Antibodies that were used for western blot analysis are: phos-TFEB (Calbiochem, ABE1971), TFEB (Bethyl, A303–673A), β-Actin (Sigma, A5441), GAPDH (Cell Signaling, 2118), p62 (Abnova, H00008878-M01), phos-s6-ribosomal protein (Cell Signaling, 4858), s6-ribosomal protein (Cell Signaling, 2217), phos-4EBP1 (Cell Signaling, 9451), 4EBP1 (Cell Signaling, 9452), phos-ERK (Cell Signaling, 9101), ERK (Cell Signaling, 9102), Lamin A/C (Cell Signaling, 2032), Lamp1 (DSHB, 1D4B), CYP2E1 (Abcam, ab28146), ALDH2 (Proteintech, 15310–1-AP). Anti-myeloperoxidase (MPO, # PP023AA) was from Biocare Medical. Antibodies for vacuolar ATPase H+ Transporting V1 Subunit A (V-ATP6V1a) and vacuolar ATPase H+ Transporting V1 Subunit B2 (V-ATP6V1b2) are kindly provided by Dr. Dennis Brown at Harvard Medical School. The Anti-LC3 antibody was generated as previously described (Ding, Li et al. 2010 (link)). HRP-conjugated secondary antibodies were from Jackson ImmunoResearch.
Chao X., Wang S., Yang L., Ni H.M, & Ding W.X. (2021). Trehalose activates hepatic transcription factor EB (TFEB) but fails to ameliorate alcohol-impaired TFEB and liver injury in mice. Alcoholism, clinical and experimental research, 45(10), 1950-1964.
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2

Antibody Characterization for Cell Biology

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The antibodies used for this study were AMYLASE (A8273; Sigma-Aldrich), Atg5 (PM050, MBL, Woburn, MA), phos-TFEB (ABE1971; Millipore, Burlington, MA), TFEB (A303-673A; Bethyl Laboratories, Montgomery, TX), CK19 (TROMA-III, Developmental Studies Hybridoma Bank, Iowa City, IA), GAPDH (2118; Cell Signaling Technology, Danvers, MA), p62 (H00008878-M01; Abnova, Taipei, Taiwan), phos-4EBP1 (9451; Cell Signaling Technology), 4EBP1 (9452; Cell Signaling Technology), phos-MAPK (9101; Cell Signaling Technology), MAPK (9102; Cell Signaling Technology), phos-S6-ribosomal protein (4858; Cell Signaling Technology), S6-ribosomal protein (2217; Cell Signaling Technology), GFP (sc-9996; Santa Cruz Biotechnology, Dallas, TX), LAMIN A/C (2032; Cell Signaling Technology), LAMP1 (1D4B and H4A3; Developmental Studies Hybridoma Bank, Iowa City, IA), LAMP2 (ABL-93 and H4A4; Developmental Studies Hybridoma Bank), and PGC1α (PAB12061; Abnova). The TAP antibody was generated by Thomas Kolodecik from Yale University.45 (link) Antibodies for VATP6V1a and VATP6V1b2 are gifts from Dr. Dennis Brown from Harvard Medical School. The Anti-LC3 antibody was generated as previously described.43 (link) HRP-conjugated, FITC-conjugated and Rhodamine-conjugated secondary antibodies were from Jackson ImmunoResearch (West Grove, PA).
Wang S., Ni H.M., Chao X., Ma X., Kolodecik T., De Lisle R., Ballabio A., Pacher P, & Ding W.X. (2020). Critical Role of TFEB-Mediated Lysosomal Biogenesis in Alcohol-Induced Pancreatitis in Mice and Humans. Cellular and Molecular Gastroenterology and Hepatology, 10(1), 59-81.
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3

Pharmacological Inhibition of mTOR Pathway

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AZD-8055 was purchased from MedChemExpress (Shanghai, China). Antibodies for western blotting against HPCAL1, ACSS2, RUVBL1, GAPDH, his tag and HA tag were obtained from Proteintech (Wuhan, China). Antibody against phos-mTOR, and phos-4EBP1 and phos-4EBP1 were purchased from Cell Signaling Technology (USA). SCD1 antibody was obtained from ABclonal (Wuhan, China). HPCAL1 antibody for IHC staining and anti-FLAG M2 gel was purchased form Sigma-Aldrich (USA).
Chen T., Yuan Z., Lei Z., Duan J., Xue J., Lu T., Yan G., Zhang L., Liu Y., Li Q, & Zhang Y. (2022). Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling. Theranostics, 12(17), 7450-7464.
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4

AMPK Signaling Modulation in Cancer Treatment

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Tamoxifen, Fulvestrant, methyl succinate, and spautin-1 were purchased from Sigma Aldrich. The following rabbit polyclonal Antibodies were obtained from Cell Signaling: AMPKα, AMPKα1, phos-AMPKα, phos-ACC, ACC, Phos-p70s6K, phos-s6, phos-4EBP1, cleaved PARP, cleaved caspase 3, and LC3B. ERα antibody purchased from Genetex. AMPKα2 antibody purchased from Millipore. siRNA against PRKAA2 was purchased from Dharmacon.
Daurio N.A., Tuttle S.W., Worth A.J., Song E.Y., Davis J.M., Snyder N.W., Blair I.A, & Koumenis C. (2016). AMPK activation and metabolic reprogramming by tamoxifen through estrogen receptor-independent mechanisms suggests new uses for this therapeutic modality in cancer treatment. Cancer research, 76(11), 3295-3306.
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5

Antibody Panel for Western Blot Analysis

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The following antibodies were used for western blot analysis: TFEB (Bethyl, A303-673A), β-Actin (Sigma, A5441), GAPDH (Cell Signaling, 2118), p62 (Abnova, H00008878-M01), GFP (Santa Cruz), phos-s6-ribosomal protein (Cell Signaling, 4858), phos-4EBP1 (Cell Signaling, 9451), s6-ribosomal protein (Cell Signaling, 2217), 4EBP1 (Cell Signaling, 9452), phos-ERK (Cell Signaling, 9101), ERK (Cell Signaling, 9102), anti-lymphocyte antigen B superfamily (Ly6B, AbD Serotec, MCA771G), Lamin A/C (Cell Signaling, 2032), Lamp1 (DSHB, 1D4B), TFE3 (Sigma, HPA023881), PGC1α (Abnova, PAB12061), mTOR (Cell Signaling, 2972). Antibodies for ATPase H+ Transporting V1 Subunit 1A (VATP6V1a) and ATPase H+ Transporting V1 Subunit 1B2 (VATP6V1b2) are gifts from Dr. Dennis Brown from Harvard Medical School. The Anti-LC3 antibody was generated as previously described5 (link). HRP-conjugated, FITC-conjugated and Cy3-conjugated secondary antibodies were from Jackson ImmunoResearch.
Chao X., Wang S., Zhao K., Li Y., Williams J.A., Li T., Chavan H., Krishnamurthy P., He X.C., Li L., Ballabio A., Ni H.M, & Ding W.X. (2018). Impaired TFEB-mediated Lysosome Biogenesis and Autophagy Promote Chronic Ethanol-induced Liver Injury and Steatosis in Mice. Gastroenterology, 155(3), 865-879.e12.
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