Anti-mTOR antibody [Y391] - ChIP Grade (ab32028)
- Product nameAnti-mTOR antibody [Y391] - ChIP GradeSee all mTOR primary antibodies ...
- DescriptionRabbit monoclonal [Y391] to mTOR - ChIP Grade
- Tested applicationsIHC-P, WB, ICC/IF, Flow Cyt, IP more details
- Species reactivityReacts with: Mouse, Rat, Human
A synthetic peptide corresponding to residues near the C-term of PI3K/PI4K domain of human mTOR/FRAP was used as an immunogen
- Epitopeab32028 reacts with an epitope located in the C terminal region of mTOR.
- Positive control
- Hela cell lysate, MCF7 cell lysate, human prostate carcinoma.
- General notes
Produced under U.S. Patent No. 5,675,063.
This product is available conjugated to DyLight® 488 see ab139837.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone numberY391
- Epigenetics and Nuclear Signaling
- DNA / RNA
- DNA Damage & Repair
- DNA Damage Response
- DNA Damage Recognition
Our Abpromise guarantee covers the use of ab32028 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|WB||WB: 1/1000 - 1/2000. Detects a band of approximately >250 kDa (predicted molecular weight: 289 kDa).|
|ICC/IF||ICC/IF: 1/50 - 1/100.|
|Flow Cyt||Flow Cyt: 1/50 - 1/80.|
- FunctionKinase subunit of both mTORC1 and mTORC2, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-421', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Phosphorylates MAF1 leading to attenuation of its RNA polymerase III-repressive function. mTORC2 is also activated by growth. factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. mTORC2 plays a critical role in AKT1 'Ser-473' phosphorylation, which may facilitate the phosphorylation of the activation loop of AKT1 on 'Thr-308' by PDK1 which is a prerequisite for full activation. mTORC2 regulates the phosphorylation of SGK1 at 'Ser-422'. mTORC2 also modulates the phosphorylation of PRKCA on 'Ser-657'.
- Tissue specificityExpressed in numerous tissues, with highest levels in testis.
- Sequence similaritiesBelongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 7 HEAT repeats.
Contains 1 PI3K/PI4K domain.
modificationsAutophosphorylated; when part of mTORC1 or mTORC2.
- Cellular localizationEndoplasmic reticulum membrane. Golgi apparatus membrane. Mitochondrion outer membrane. Lysosome. Cytoplasm. Nucleus > PML body. Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB.
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Anti-mTOR antibody [Y391] - ChIP Grade images
Overlay histogram showing HeLa cells stained with ab32028 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32028, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
Immunohistochemical analysis of mTOR expression in paraffin-embedded human prostate carcinoma, using 1/250 ab32028.
ICC/IF image of ab32028 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32028, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-mTOR antibody [Y391] - ChIP Grade (ab32028) at 1/2000 dilution
Lane 1 : Hela cell lysate
Lane 2 : MCF-7
Predicted band size : 289 kDa
Observed band size : >250 kDa (why is the actual band size different from the predicted?)
References for Anti-mTOR antibody [Y391] - ChIP Grade (ab32028)
This product has been referenced in:
- Fleming GF et al. Phase II trial of temsirolimus in patients with metastatic breast cancer. Breast Cancer Res Treat 136:355-63 (2012). Read more (PubMed: 22245973) »
- Gant JC et al. Disrupting function of FK506-binding protein 1b/12.6 induces the Ca²+-dysregulation aging phenotype in hippocampal neurons. J Neurosci 31:1693-703 (2011). Rat . Read more (PubMed: 21289178) »