Anti-mTOR 抗体 (ab2732)
Key features and details
- Rabbit polyclonal to mTOR
- Suitable for: ICC/IF, IP, WB
- Reacts with: Rat, Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
-
製品名
Anti-mTOR antibody
mTOR 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to mTOR -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, IP, WBmore details -
種交差性
交差種: Rat, Human
交差が予測される動物種: Mouse
-
免疫原
-
ポジティブ･コントロール
- ICC/IF: HepG2 cells. L6 myotubes. IP: HeLa lysates. WB: HeLa, HEK-293T, Jurkat-HepG2 and LNCaP whole cell lysate.
-
特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
バッファー
pH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
Concentration information loading... -
特記事項（精製）
Affinity purified using the immunising peptideimmobilized on solid support. -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab2732の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| ICC/IF | (1) |
1/100.
|
| IP |
Use at 2-10 µg/mg of lysate.
|
|
| WB | (7) |
1/2000 - 1/10000. Predicted molecular weight: 289 kDa.
|
| 特記事項 |
|---|
|
ICC/IF
1/100. |
|
IP
Use at 2-10 µg/mg of lysate. |
|
WB
1/2000 - 1/10000. Predicted molecular weight: 289 kDa. |
ターゲット情報
-
機能
Kinase 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'. -
組織特異性
Expressed in numerous tissues, with highest levels in testis. -
配列類似性
Belongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 7 HEAT repeats.
Contains 1 PI3K/PI4K domain. -
翻訳後修飾
Autophosphorylated; when part of mTORC1 or mTORC2. -
細胞内局在
Endoplasmic 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
-
参照データベース
- Entrez Gene: 2475 Human
- Entrez Gene: 56717 Mouse
- Entrez Gene: 56718 Rat
- Omim: 601231 Human
- SwissProt: P42345 Human
- SwissProt: Q9JLN9 Mouse
- SwissProt: P42346 Rat
- Unigene: 338207 Human
see all -
別名
- 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
see all
画像
-
Western blot - Anti-mTOR antibody (ab2732)All lanes : Anti-mTOR antibody (ab2732) at 1/2000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : MTOR [homo] CRISPR-Cas9 edited A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 289 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-mTOR antibody staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab2732 was shown to bind specifically to mTOR. A band was observed at 250 kDa in wild-type A549 cell lysates with no signal observed at this size in MTOR CRISPR-Cas9 edited cell line ab283257. The band observed in the CRISPR-Cas9 edited lysate lane below 250 kDa is likely to represent a truncated form of mTOR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MTOR CRISPR-Cas9 edited A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
Western blot - Anti-mTOR antibody (ab2732)All lanes : Anti-mTOR antibody (ab2732) at 0.1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 5 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 50 µg per lane.
Predicted band size: 289 kDa
Exposure time: 3 minutesLysates prepared using NETN lysis buffer.
-
Immunoprecipitation - Anti-mTOR antibody (ab2732)mTOR was immunoprecipitated from HeLa cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab2732 at 3 µg per reaction. mTOR was also immunoprecipitated by ab2833. Western blot was performed from the immunoprecipitate with ab2732 at 1 µg/ml.
Lane 1: ab2833 IP in HeLa whole cell lysate.
Lane 2: ab2732 IP in HeLa whole cell lysate.
Lane 3: Control IgG IP in HeLa whole cell lysate.Detection: Chemiluminescence
-
Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)ICC/IF image of ab2732 stained HepG2 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 (ab2732, 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. -
Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)ab2732 at a 1:100 dilution confocally staining mTOR (red) in L6 myotubes, alongside a nuclear antigen antibody (green).
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (349)
ab2732 は 349 報の論文で使用されています。
- Lu Y et al. Impairment of Autophagy Mediates the Uric-Acid-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells. Pharmacology 109:34-42 (2024). PubMed: 38011839
- Ning S et al. MicroRNA-494 regulates high glucose-induced cardiomyocyte apoptosis and autophagy by PI3K/AKT/mTOR signalling pathway. ESC Heart Fail 10:1401-1411 (2023). PubMed: 36772911
- Wang Y et al. Rumen-protected glucose stimulates the secretion of reproductive hormones and the mTOR/AKT signaling pathway in the ovaries of early postpartum. Sci Rep 13:2940 (2023). PubMed: 36808140
- Zhu S et al. DNA methylation and miR-92a-3p-mediated repression of HIP1R promotes pancreatic cancer progression by activating the PI3K/AKT pathway. J Cell Mol Med 27:788-802 (2023). PubMed: 36811277
- Ibrahim YH et al. An affinity tool for the isolation of endogenous active mTORC1 from various cellular sources. J Biol Chem 299:104644 (2023). PubMed: 36965617