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Deptor

Manufactured by Novus Biologicals
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DEPTOR is a protein that functions as a negative regulator of the mTOR (mammalian target of rapamycin) signaling pathway. It inhibits the activity of both mTOR complexes, mTORC1 and mTORC2, thereby modulating cellular processes such as cell growth, proliferation, and metabolism.

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Lab products found in correlation

Axiovert 200m, by Zeiss (4 mentions) Ecl plus developing system, by Cytiva (2 mentions) P473akt, by Cell Signaling Technology (2 mentions) 4e bp1, by Cell Signaling Technology (2 mentions) Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Tritc conjugated antibodies, by Jackson ImmunoResearch (2 mentions) P erk, by Cell Signaling Technology (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Vectastain elite abc hrp kit, by Vector Laboratories (1 mentions) Flag m2, by Merck Group (1 mentions) 1 2 dioctanoyl sn glycero 3 pa, by Avanti Polar Lipids (1 mentions)

6 protocols using deptor

1

Western Blot Analysis of Neural Markers

Cited 1 time
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Cell lysates were created and Western blotting conducted as described (24 (link)), blocking without detergent. Membranes were probed with antibodies for MAF (Imgenex), SOX9 (Santa Cruz), S100β (Dako), MBP (Chemicon), BLBP (Millipore), DEPTOR (Novus), cleaved-caspase 3, pS6, S6, p4E-BP1, 4E-BP1, p473AKT, AKT, and β-ACTIN (Cell Signaling) as a loading control. Horseradish peroxidase-conjugated secondary antibodies (BioRad) were used with ECL Plus developing system (Amersham Biosciences).
Brundage M.E., Tandon P., Eaves D.W., Williams J.P., Miller S.J., Hennigan R.H., Jegga A., Cripe T.P, & Ratner N. (2014). MAF mediates crosstalk between Ras-MAPK and mTOR signaling in NF1. Oncogene, 33(49), 5626-5636.
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2

Immunofluorescence and Immunohistochemistry Protocol

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Cultured cells were fixed, permeabilized with 0.2% Triton-X 100 and blocked with 10% normal serum for 1h at room temperature (19 (link)). Primary antibodies used were S100β (Dako), MBP (Chemicon), and BLBP (Millipore). Secondary incubations used host-appropriate TRITC conjugated antibodies (Jackson Immunoresearch). Nuclei were labeled with DAPI (Sigma-Aldrich) and microscopic images acquired with ImageJ software (NIH) on a Zeiss Axiovert 200M. Tumor paraffin sections stained for MAF (Imgenex), pS6, pERK (Cell Signaling), and DEPTOR (Novus) were visualized with DAB.
Brundage M.E., Tandon P., Eaves D.W., Williams J.P., Miller S.J., Hennigan R.H., Jegga A., Cripe T.P, & Ratner N. (2014). MAF mediates crosstalk between Ras-MAPK and mTOR signaling in NF1. Oncogene, 33(49), 5626-5636.
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3

Immunofluorescence and Immunohistochemistry Protocol

Cited 1 time
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Cultured cells were fixed, permeabilized with 0.2% Triton-X 100 and blocked with 10% normal serum for 1h at room temperature (19 (link)). Primary antibodies used were S100β (Dako), MBP (Chemicon), and BLBP (Millipore). Secondary incubations used host-appropriate TRITC conjugated antibodies (Jackson Immunoresearch). Nuclei were labeled with DAPI (Sigma-Aldrich) and microscopic images acquired with ImageJ software (NIH) on a Zeiss Axiovert 200M. Tumor paraffin sections stained for MAF (Imgenex), pS6, pERK (Cell Signaling), and DEPTOR (Novus) were visualized with DAB.
Brundage M.E., Tandon P., Eaves D.W., Williams J.P., Miller S.J., Hennigan R.H., Jegga A., Cripe T.P, & Ratner N. (2014). MAF mediates crosstalk between Ras-MAPK and mTOR signaling in NF1. Oncogene, 33(49), 5626-5636.
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4

Phosphorylated p70 and DEPTOR Immunohistochemistry

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Four um-thick sections of tissue blocks were cut and stained using the Vectastain Elite ABC HRP kit (Vector, Burlingham, CA) in a two-step IHC technique as previously described (8 , 9 (link)). Antigen retrieval was performed and slides were incubated with primary antibody against phosphorylated p70 (1:500 dilution) from InVitrogen (Carlsbad, CA) or DEPTOR (1:750 dilution) from Novus (Littleton, CO).
Vega M.I., Shi Y., Frost P., Huerta-Yepez S., Antonio-Andres G., Hernandez-Pando R., Lee J., Jung M.E., Gera J.F, & Lichtenstein A. (2019). A Novel Therapeutic Induces DEPTOR Degradation in Multiple Myeloma Cells with Resulting Tumor Cytotoxicity. Molecular cancer therapeutics, 18(10), 1822-1831.
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5

Antibody Source Identification

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Antibodies were obtained from the following sources: FLAG M2 from Sigma-Aldrich (St. Louis, MO); DEPTOR from Novus; tubulin from Abcam (Cambridge, UK); raptor, rictor, and mSin1 from Bethyl laboratory (Montgomery, TX); mTOR from Santa Cruz Biotechnology (Santa Cruz, CA); and all other antibodies from Cell Signaling Technology (Danvers, MA). All secondary antibodies were from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA). 1,2-Dioctanoyl-sn-glycero-3–PA was obtained from Avanti Polar Lipids (Alabaster, AL). All other reagents were from Sigma-Aldrich (St. Louis, MO).
Baek M.O., Song H.I., Han J.S, & Yoon M.S. (2018). Differential regulation of mTORC1 and mTORC2 is critical for 8-Br-cAMP-induced decidualization. Experimental & Molecular Medicine, 50(10), 1-11.
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6

Western Blot Analysis of Neural Markers

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were created and Western blotting conducted as described (24 (link)), blocking without detergent. Membranes were probed with antibodies for MAF (Imgenex), SOX9 (Santa Cruz), S100β (Dako), MBP (Chemicon), BLBP (Millipore), DEPTOR (Novus), cleaved-caspase 3, pS6, S6, p4E-BP1, 4E-BP1, p473AKT, AKT, and β-ACTIN (Cell Signaling) as a loading control. Horseradish peroxidase-conjugated secondary antibodies (BioRad) were used with ECL Plus developing system (Amersham Biosciences).
Brundage M.E., Tandon P., Eaves D.W., Williams J.P., Miller S.J., Hennigan R.H., Jegga A., Cripe T.P, & Ratner N. (2014). MAF mediates crosstalk between Ras-MAPK and mTOR signaling in NF1. Oncogene, 33(49), 5626-5636.
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