Product nameHuman MTOR knockout HEK293T cell lysate
See all mTOR kits
Cell line information
Parental cell line: HEK293T
Organism: Homo sapiens
Gene editing information
Editing tool: CRISPR/Cas9
Mutation: Homozygous: 180 bp insertion in exon 2.
Knockout validation: Confirmed by Sanger sequencing and WB.
Reconstitution instructions: To use as a WB control, resuspend in 45 µL of Sample buffer (40% (w/v) Glycerol, 4% (w/v) Lithium Dodecyl Sulfate, 4% Ficoll 400, 0.025% Phenol Red, 0.025% Brilliant Blue G250, 2 mM EDTA) and 5 µL of DTT to resuspend @ 2mg/ml. Mix well, then boil the sample for 10 minutes before loading it onto the gel.
User storage instructions: Upon receiving, lysate can be diluted with 1 x SDS sample buffer & will be stable at -20°C for 12 months. Long term storage at -80°C.
Knockout validationSanger Sequencing, Western Blot (WB)
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit Human MTOR knockout HEK293T cell lysate (Lyophilized) 1 x 100µg Human Wild Type HEK293T cell lysate (Lyophilized) 1 x 100µg
- Epigenetics and Nuclear Signaling
- DNA / RNA
- DNA Damage & Repair
- DNA Damage Response
- DNA Damage Recognition
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.
- Information by UniProt
- dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1)
- FK506 binding protein 12 rapamycin associated protein 1
- FK506 binding protein 12 rapamycin associated protein 2
Our Abpromise guarantee covers the use of ab263789 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.|
Lane 1: HEK-293 wildtype cell lysate (20 µg)
Lane 2: MTOR HEK-293 knockout cell lysate (20 µg)
Lane 3: Wild-type HEK-293T cell lysate (20 µg)
Lane 4: MTOR knockout HEK-293T cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab134903 observed at 289 kDa. Red - loading control, ab130007 observed at 125 kDa.
ab134903 was shown to react with mTOR in wild-type HEK-293T cells. Loss of signal was observed when knockout sample ab263789 was used. Wild-type and mTOR knockout samples were subjected to SDS-PAGE. ab134903 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Homozygous: 180 bp deletion in exon 2
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab263789 has not yet been referenced specifically in any publications.