Anti-RICTOR 抗体 (ab105469)
Key features and details
- Rabbit polyclonal to RICTOR
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, 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
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製品名
Anti-RICTOR antibody
RICTOR 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to RICTOR -
由来種
Rabbit -
アプリケーション
適用あり: WB, ICC/IFmore details -
種交差性
交差種: Mouse, Human
交差が予測される動物種: Rat, Rabbit, Horse, Cow, Dog, Pig, Macaque monkey, Gorilla, Chinese hamster
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免疫原
Synthetic peptide corresponding to Human RICTOR aa 250-350 conjugated to keyhole limpet haemocyanin.
(Peptide available asab124137) -
ポジティブ･コントロール
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; Jurkat; HepG2; NIH3T3.
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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
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
精製度
Immunogen affinity purified -
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
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Compatible Secondaries
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Isotype control
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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ab105469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 |
|---|---|---|
| WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).
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| ICC/IF |
Use a concentration of 5 µg/ml.
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| 特記事項 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa). |
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ICC/IF
Use a concentration of 5 µg/ml. |
ターゲット情報
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機能
Subunit of mTORC2, which regulates cell growth and survival in response to hormonal signals. mTORC2 is activated by growth factors, but, in contrast to mTORC1, 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'. Plays an essential role in embryonic growth and development. -
配列類似性
Belongs to the RICTOR family. -
翻訳後修飾
Phosphorylated by MTOR; when part of mTORC2. Phosphorylated at Thr-1135 by RPS6KB1; phosphorylation of RICTOR inhibits mTORC2 and AKT1 signaling. - Information by UniProt
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参照データベース
- Entrez Gene: 479356 Dog
- Entrez Gene: 101132782 Gorilla
- Entrez Gene: 253260 Human
- Entrez Gene: 78757 Mouse
- Entrez Gene: 310131 Rat
- Omim: 609022 Human
- SwissProt: Q6R327 Human
- SwissProt: Q6QI06 Mouse
see all -
別名
- AVO3 antibody
- AVO3 homolog antibody
- DKFZp686B11164 antibody
see all
画像
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Western blot - Anti-RICTOR antibody (ab105469)All lanes : Anti-RICTOR antibody (ab105469) at 1 µg/ml
Lane 1 : Wild-type A549 cell lysate
Lane 2 : RICTOR knockout A549 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 192 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-RICTOR antibody staining at 1 ug/ml, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab105469 was shown to bind specifically to RICTOR. A band was observed at 190 kDa in wild-type A549 cell lysates with no signal observed at this size in RICTOR knockout cell line ab277866 (knockout cell lysate ab288315). To generate this image, wild-type and RICTOR knockout 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Western blot - Anti-RICTOR antibody (ab105469)Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: RICTOR knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab105469 observed at 192 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab105469 was shown to recognize RICTOR in wild-type HAP1 cells as signal was lost at the expected MW in RICTOR knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and RICTOR knockout samples were subjected to SDS-PAGE. ab105469 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 µg/mL and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-RICTOR antibody (ab105469)All lanes : Anti-RICTOR antibody (ab105469) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 192 kDa
Observed band size: 192 kDa
Additional bands at: 37 kDa, 52 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Immunocytochemistry/ Immunofluorescence - Anti-RICTOR antibody (ab105469)ICC/IF image of ab105469 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 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 (ab105469, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) MCF7 cells at 5µg/ml.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (0)
ab105469 は論文での使用が確認できていません。