Anti-mTOR抗体
参阅全部 mTOR 一抗
兔多克隆抗体to mTOR
Rabbit
适用于: ICC/IF, IP, WBmore details
与反应: Rat, Human
预测可用于: Mouse
Synthetic peptide within Human mTOR aa 200-250. The exact sequence is proprietary.
Database link: P42345
(Peptide available as ab39393)
ICC/IF: HepG2 cells. L6 myotubes. IP: HeLa lysates. WB: HeLa, HEK-293T, Jurkat-HepG2 and LNCaP whole cell lysate.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
pH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
浓度
50 µg 浓度为 1 mg/ml
Affinity purified using the immunising peptideimmobilized on solid support.
多克隆
IgG
Abpromise™承诺保证使用ab2732于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| 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. |
Entrez Gene: 2475 Human
Entrez Gene: 56717 Mouse
Omim: 601231 Human
SwissProt: P42345 Human
SwissProt: Q9JLN9 Mouse
Unigene: 338207 Human
Unigene: 21158 Mouse
Unigene: 11008 Rat
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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 kDawhy 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 minutes
Lysates 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).
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