Anti-RICTOR antibody (ab105469)
Key features and details
- Rabbit polyclonal to RICTOR
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-RICTOR antibody
See all RICTOR primary antibodies -
Description
Rabbit polyclonal to RICTOR -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human RICTOR aa 250-350 conjugated to keyhole limpet haemocyanin.
(Peptide available asab124137) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; Jurkat; HepG2; NIH3T3.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab105469 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications RatRabbitHorseCowDogPigMacaque monkeyGorillaChinese hamsterApplication Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).ICC/IF Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Detects a band of approximately 192 kDa (predicted molecular weight: 192 kDa).ICC/IF
Use a concentration of 5 µg/ml.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: 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 -
Alternative names
- AVO3 antibody
- AVO3 homolog antibody
- DKFZp686B11164 antibody
see all
Images
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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.
Protocols
References (0)
ab105469 has not yet been referenced specifically in any publications.
Images
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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.
