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mTOR (human FRB Domain) polyclonal antibody

ALX-215-065-1 1 Vial 586.00 USD
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Product Details

Alternative Name:mTOR (human FKBP12-rapamycin-binding domain)
Immunogen:Recombinant human mTOR (FKBP12-rapamycin-binding (FRB) domain).
UniProt ID:P42345
Species reactivity:Human
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.
Applications:ICC, WB
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.
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).
Formulation:Lyophilized from neat serum containing sodium azide.
Reconstitution:Reconstitute with 50µl water.
Shipping:Blue Ice
Long Term Storage:+4°C
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).
Regulatory Status:RUO - Research Use Only
mTOR (human FRB Domain) polyclonal antibody Western blot
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.
mTOR (human FRB Domain) polyclonal antibody WB
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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mTOR (human FRB Domain) polyclonal antibody Western blot mTOR (human FRB Domain) polyclonal antibody WB

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;

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