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mTOR (FKBP12-rapamycin associated protein (FRAP)) is one of a family of proteins involved in cell cycle progression, DNA recombination, and DNA damage detection. It has been shown to associate with the immunophilin FKBP12 in a rapamycin-dependent fashion. FKBP12-rapamycin-binding (FRB) domain is a conserved 11kDa region necessary for FKBP12-rapamycin binding. FRB is one of several functional domains of mTOR: HEAT (Huntington elongation factor 1A-protein phosphatase 2A-A subunit-TOR); FAT (FRAP, ATM, TTRAP2); PIKK (PI3-kinase-related kinase); RD (regulatory domain); and FATC (FAT, C terminal).
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Figure: WB using PAb to mTOR (human FRB Domain) (Prod. No. ALX-215-065) on total yeast extracts derived from cells expressing FRB-tagged genes coding for three different transcription factors. Separation SDS-PAGE.”””,mTOR (human FRB Domain) polyclonal antibody WB,mTOR (human FRB Domain) polyclonal antibody Western Blot,yes
ALX-215-065,215-065_2_WB.gif,http://static.enzolifesciences.com/fileadmin/files/image/215-065_2_WB.gif,Figure 2: Western Blot of mTOR (human FRB Domain)
Figure 2: Western Blot of mTOR (human FRB Domain), pAb (Prod. No. ALX-215-065) with Hela cells.
Method:Two ml of an exponential HeLa suspension culture (2-4×105 cells) was washed twice with PBS quickly without resuspention of the pellet by spinning in a Eppendorf tube and finally with 25% TCA. TCA was carefully removed and pellet respuspended 100µl FSB + ßME (final sample buffer) and the pH adjusted with about a few µl of 1M Tris base (≈10 µl). The sample was heated at 100°C for 5 min. and sonicated. SDS PAGE was at 7.5%, 20 & 25 µl loaded in lane 1 and 2, respectively. The TCA step is performed to prevent proteolytic breakdown of the very large mTOR.
| Regulatory Status |
RUO – Research Use Only |
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Last modified: January 23, 2025