WB
Western blot analysis of mTOR on different lysates with Rabbit anti-mTOR antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: MCF7 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: HepG2 cell lysate (20 µg/Lane), Lane 5: C2C12 cell lysate (20 µg/Lane), Lane 6: PC-12 cell lysate (20 µg/Lane), Lane 7: C6 cell lysate (20 µg/Lane), Lane 8: Mouse testis tissue lysate (20 µg/Lane), Lane 9: Rat testis tissue lysate (20 µg/Lane), Exposure time: 24 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-mTOR antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-mTOR antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling mTOR with Rabbit anti-mTOR antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-mTOR antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling mTOR with Rabbit anti-mTOR antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-mTOR antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | mTOR Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The phosphatidylinositol kinase (PIK) family members fall into two distinct subgroups. The first subgroup contains proteins such as the PI 3- and PI 4-kinases and the second group comprises the PIK-related kinases. The PIK-related kinases include Atm, DNA-PKCS and FRAP. These proteins have in common a region of homology at their carboxy-termini that is not present in the PI 3- and PI 4-kinases. The Atm gene is mutated in the autosomal recessive disorder ataxia telangiectasia (AT) that is characterized by cerebellar degeneration (ataxia) and the appearance of dilated blood vessels (telangiectases) in the conjunctivae of the eyes. AT cells are hypersensitive to ionizing radiation, impaired in mediating the inhibition of DNA synthesis and display delays in p53 induction. DNA-PK is a heterotrimeric DNA binding enzyme that is composed of a large subunit, DNA-PKCS, and two smaller subunits collectively known as Ku. The loss of DNA-PK leads to defects in DSB repair and V(D)J recombination. FRAP can autophosphorylate on serine and bind to Rapamycin/FKBP. FRAP is also an upstream regulator of S6 kinase and has been implicated in the regulation of p27 and p21 expression. |
| Immunogen | Synthetic peptide within human mTOR aa 2400-2440. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:5000 IHC:1:200-1:1000 ICC/IF:1:50-1:200 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) antibody
FK506 binding protein 12 rapamycin associated protein 1 antibody
FK506 binding protein 12 rapamycin associated protein 2 antibody FK506 binding protein 12 rapamycin complex associated protein 1 antibody FK506-binding protein 12-rapamycin complex-associated protein 1 antibody FKBP rapamycin associated protein antibody FKBP12 rapamycin complex associated protein antibody FKBP12-rapamycin complex-associated protein 1 antibody FKBP12-rapamycin complex-associated protein antibody FLJ44809 antibody FRAP antibody FRAP1 antibody FRAP2 antibody Mammalian target of rapamycin antibody Mechanistic target of rapamycin antibody mTOR antibody MTOR_HUMAN antibody OTTHUMP00000001983 antibody RAFT1 antibody Rapamycin and FKBP12 target 1 antibody Rapamycin associated protein FRAP2 antibody Rapamycin target protein 1 antibody Rapamycin target protein antibody RAPT1 antibody Serine/threonine-protein kinase mTOR antibody |
|---|---|
| Molecular Weight(MW) | 289kDa |
| Cellular Localization | Endoplasmic reticulum membrane, Microsome membrane, PML body, Golgi apparatus membrane, Cytoplasm, Lysosome, Lysosome membrane, Mitochondrion outer membrane, phagosome. |
WB
Western blot analysis of mTOR on different lysates with Rabbit anti-mTOR antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: MCF7 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: HepG2 cell lysate (20 µg/Lane), Lane 5: C2C12 cell lysate (20 µg/Lane), Lane 6: PC-12 cell lysate (20 µg/Lane), Lane 7: C6 cell lysate (20 µg/Lane), Lane 8: Mouse testis tissue lysate (20 µg/Lane), Lane 9: Rat testis tissue lysate (20 µg/Lane), Exposure time: 24 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-mTOR antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-mTOR antibody at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling mTOR with Rabbit anti-mTOR antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-mTOR antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling mTOR with Rabbit anti-mTOR antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-mTOR antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.| Application Notes | WB:1:5000 IHC:1:200-1:1000 ICC/IF:1:50-1:200 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
