Recombinant Anti-RICTOR antibody [EPR22008] (ab219950)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22008] to RICTOR
- Suitable for: Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RICTOR antibody [EPR22008]
See all RICTOR primary antibodies -
Description
Rabbit monoclonal [EPR22008] to RICTOR -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, HEK-293, HCT 116, MDA-MB-231, SW480 and PC-3 whole cell lysate. Flow Cyt (intra): HCT 116 and HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22008 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab219950 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 192 kDa).
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 192 kDa). |
Target
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Function
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. -
Sequence similarities
Belongs to the RICTOR family. -
Post-translational
modificationsPhosphorylated 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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Database links
- Entrez Gene: 253260 Human
- Omim: 609022 Human
- SwissProt: Q6R327 Human
- Unigene: 407926 Human
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Alternative names
- AVO3 antibody
- AVO3 homolog antibody
- DKFZp686B11164 antibody
see all
Images
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All lanes : Anti-RICTOR antibody [EPR22008] (ab219950) at 1/1000 dilution
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 [EPR22008] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab219950 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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All lanes : Anti-RICTOR antibody [EPR22008] (ab219950) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate at 20 µg
Lane 2 : RICTOR knockout HAP1 whole cell lysate at 20 µg
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 4 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 192 kDaBlocking/Diluting buffer and concentration: 5%NFDM/TBST.
Exposure time: 26 seconds.
ab219950 was shown to specifically react with RICTOR in wild-type HAP1 cells as signal was lost in RICTOR knockout cells. Wild-type and RICTOR knockout samples were subjected to SDS-PAGE. ab219950 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
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All lanes : Anti-RICTOR antibody [EPR22008] (ab219950) at 1/1000 dilution
Lane 1 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lane 2 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 4 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 192 kDaBlocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: Lane 1: 3 minutes; Lane 2: 48 seconds; Lane 3-4: 26 seconds.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling RICTOR with ab219950 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HCT 116 (human colorectal carcinoma cell line) cell line labeling RICTOR with ab219950 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab219950 has not yet been referenced specifically in any publications.