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mTOR (human FRB Domain) polyclonal antibody

 
ALX-215-065-1 1 Vial 443.00 USD
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Product Specification

Alternative Name:mTOR (human FKBP12-rapamycin-binding domain)
 
Host:Rabbit
 
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
ChIP
 
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:Shipped on 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).
 
215-065
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.
215-065 2 WB
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.
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215-065 215-065 2 WB

Product Literature References

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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