Product Details
| Alternative Name: | mTOR (human FKBP12-rapamycin-binding domain) |
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| Host: | Rabbit |
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| Immunogen: | Recombinant human mTOR (FKBP12-rapamycin-binding (FRB) domain). |
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| UniProt ID: | P42345 |
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| Species reactivity: | Human
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| Specificity: | Recognizes the FRB domain of fusion proteins expressed in Saccharomyces cerevisiae (see H. Haruki, et al. 2008) and the human FRB domain of mTOR. |
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| Applications: | ICC, WB
ChIP
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| Recommended Dilutions/Conditions: | Western Blot (1:2,000)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application. |
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| Application Notes: | Among other applications this antibody can be used to control FRB-tagging of genes and the level of expression thereof in yeast cells (see H. Haruki, et al. 2008). |
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| Formulation: | Lyophilized from neat serum containing sodium azide. |
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| Reconstitution: | Reconstitute with 50µl water. |
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| Shipping: | Blue Ice |
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| Long Term Storage: | +4°C |
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| Scientific Background: | 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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| Regulatory Status: | RUO - Research Use Only |
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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 (2-4x105 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.
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."
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Product Literature References
DiCre-Based inducible gene expression: J.D. Damasceno, et al.; Methods Mol. Biol.
1971, 225 (2019),
Abstract;
Autoinhibitory regulation of S100A8/S100A9 alarmin
activity locally restricts sterile inflammation: T. Vogl, et al.; J. Clin. Invest.
128, 1852 (2018),
Application(s): Immunofluorescence on human alveolar epithelial cells,
Abstract;
Full Text
Genome-wide Analysis of RNA Polymerase II Termination at Protein-Coding Genes: C. Baejen, et al.; Mol. Cell
66, 38 (2017),
Abstract;
Evidence for Multiple Mediator Complexes in Yeast Independently Recruited by Activated Heat Shock Factor: J. Anandhakumar, et al.; Mol. Cell. Biol.
36, 1943 (2016),
Application(s): ChIP from yeast cultures,
Abstract;
Full Text
Kin28 regulates the transient association of Mediator with core promoters: C. Jeronimo & F. Robert; Nat. Struct. Mol. Biol.
21, 449 (2014),
Application(s): ChIP from yeast cultures,
Abstract;
Full Text
Conditional genome engineering in Toxoplasma gondii uncovers alternative invasion mechanisms: N. Andenmatten, et al.; Nat. Methods
10, 125 (2013),
Application(s): WB in T. gondii samples,
Abstract;
Full Text
Organelle-specific, rapid induction of molecular activities and membrane tethering: T. Komatsu, et al.; Nat. Methods
7, 206 (2010),
Abstract;
Full Text
The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes: H. Haruki, et al.; Mol. Cell
31, 925 (2008),
Abstract;
