Anti-RICTOR antibody [7B3] (ab104838)

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Key features and details

  • Mouse monoclonal [7B3] to RICTOR
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Mouse, Human, Monkey
  • Isotype: IgG1

Overview

  • Product name

    Anti-RICTOR antibody [7B3]
    See all RICTOR primary antibodies

  • Description

    Mouse monoclonal [7B3] to RICTOR

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human, Monkey

  • Immunogen

    Purified recombinant fragment of Human RICTOR expressed in E. Coli.

  • Positive control

    • HeLa, PANC-1, MOLT4, HepG2 and HEK293 cell lysates. Thyroid gland tissues and Human breast carcinoma. NIH3T3 cells.

  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR173148-16 are from Tissue Culture Supernatant

Properties

Applications

Our Abpromise guarantee covers the use of ab104838 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
WB 1/500 - 1/2000. Predicted molecular weight: 192 kDa.
IHC-P 1/200 - 1/1000.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF 1/200 - 1/1000.

Target

  • 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
    modifications

    Phosphorylated by MTOR; when part of mTORC2. Phosphorylated at Thr-1135 by RPS6KB1; phosphorylation of RICTOR inhibits mTORC2 and AKT1 signaling.
  • Information by UniProt

  • Database links

    • Alternative names

      • AVO3 antibody
      • AVO3 homolog antibody
      • DKFZp686B11164 antibody
      • hAVO3 antibody
      • KIAA1999 antibody
      • Likely ortholog of mouse TORC2 specific protein AVO3 (S. cerevisiae) antibody
      • mAVO3 antibody
      • MGC39830 antibody
      • PIA antibody
      • Pianissimo antibody
      • Rapamycin insensitive companion of mTOR antibody
      • Rapamycin-insensitive companion of mTOR antibody
      • Rictor antibody
      • RICTR antibody
      • RICTR_HUMAN antibody
      • RPTOR independent companion of MTOR complex 2 antibody
      • TORC2 specific protein AVO3 antibody
      see all

    Images

    • Western blot - Anti-RICTOR antibody [7B3] (ab104838)
      Western blot - Anti-RICTOR antibody [7B3] (ab104838)
      All lanes : Anti-RICTOR antibody [7B3] (ab104838) at 1/500 dilution

      Lane 1 : HeLa cell lysate.
      Lane 2 : PANC-1 cell lysate.
      Lane 3 : MOLT4 cell lysate.
      Lane 4 : HepG2 cell lysate.
      Lane 5 : HEK293 cell lysate.

      Predicted band size: 192 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-RICTOR antibody [7B3] (ab104838)
      Immunocytochemistry/ Immunofluorescence - Anti-RICTOR antibody [7B3] (ab104838)
      ab104838 at 1/200 dilution staining RICTOR in NIH3T3 cells (green) by immunofluorescence. DRAQ5 fluorescent DNA dye is shown in blue. Actin filaments have been labeled with Alexa Fluor-555 phalloidin (red).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RICTOR antibody [7B3] (ab104838)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RICTOR antibody [7B3] (ab104838)
      ab104838 at 1/200 dilution staining RICTOR in paraffin-embedded thyroid gland tissues by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RICTOR antibody [7B3] (ab104838)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RICTOR antibody [7B3] (ab104838)
      ab104838 at 1/200 dilution staining RICTOR in paraffin-fixed Human breast carcinoma by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    • Flow Cytometry - Anti-RICTOR antibody [7B3] (ab104838)
      Flow Cytometry - Anti-RICTOR antibody [7B3] (ab104838)
      ab104838 used for Flow cytometric analysis of HeLa cells (green). The negative control is shown in purple.

    References

    Publishing research using ab104838? Please let us know so that we can cite the reference in this datasheet.

    ab104838 has been referenced in 3 publications.

    • Zhang E  et al. Minocycline promotes cardiomyocyte mitochondrial autophagy and cardiomyocyte autophagy to prevent sepsis-induced cardiac dysfunction by Akt/mTOR signaling. Apoptosis 24:369-381 (2019). PubMed: 30756206
    • Frith JE  et al. Mechanically-sensitive miRNAs bias human mesenchymal stem cell fate via mTOR signalling. Nat Commun 9:257 (2018). PubMed: 29343687
    • Hütt-Cabezas M  et al. Activation of mTORC1/mTORC2 signaling in pediatric low-grade glioma and pilocytic astrocytoma reveals mTOR as a therapeutic target. Neuro Oncol 15:1604-14 (2013). IHC-P ; Human . PubMed: 24203892

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    Dear ,
    please find attach the filled questionnaire.
    thank you very much

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    Answer

    Thank you very much for taking the time to complete and send the questionnaire.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality.

    Unfortunately none of these antibodies have been tested in frozen sections, and therefore they are not guaranteed in this application, nor do we have specific protocols tips to provide for it.

    However, there are some suggestions that may optimise the antibody performance, if you wish to give them a try:

    -I would recommend decreasing the permeabilization conditions. Both proteins are cytoplasmic; there is no need for such an aggressive permeabilization step. I would repeat the staining skipping this step and removing Triton from the blocking and the washing buffer as well as from the antibody diluent.

    -Use milder blocking conditions, using 5% of serum if possible from the same species as the secondary was raised in, to avoid cross reactivity.

    -Try using more concentrated dilutions, especially for Raptor antibody (1/50 – 1/25).

    If possible I would recommend trying both antibodies in paraffin embedded sections, as they are both guaranteed for it. The link to our protocols page is below, where you can find all the information regarding the different applications protocols, and very useful information for performing paraffin embedded immunostaining.

    https://www.abcam.com/index.html?pageconfig=popular_protocols

    Should the suggestions not improve the results, please do not hesitate to contact me again and I’ll be more than happy to help you further.

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    Question

    Dear Sr./ Mrs.,
    I am writing to you to discuss a problem we have just had with two
    antibodies purchased at your company, Rictor (AB104838) and Raptor
    (AB40768).
    We have just performed immunofluorescence staining on mouse cerebral
    tissue The staining did not work out and was similar to a negative
    control (without primary antibody and plus secondary antibody).
    The protocol that we used was the following:
    Sections were washed twice with PBS and incubated with NH4Cl 50 mM
    during 30 min. Tissue was permeated along 20 min by treatment with PBS
    containing 0.5% Triton X-100 and blocked during 1 h at room
    temperature in PBS plus 0.2% bovine serum albumin, 0.2% glycine, 0.2%
    sodium azide, 0.5% Triton X-100 and 5% normal goat serum. Slices were
    incubated overnight at 4°C with the corresponding primary antibodies
    in a buffer containing PBS plus 0.3% Triton X-100, 0.2% bovine serum
    albumin and 0.2% sodium azide. After primary antibody incubation,
    slices were washed in PBS twice for 10 min and incubated 2 h at room
    temperature with subtype-specific fluorescent secondary antibodies.
    After two consecutive 10 min-washes, slices were mounted with Mowiol
    on silane-coated slides.
    Primary antibodies were diluted 1:200 in the case of Rictor, and and
    1:100 in the case of Raptor, as recommended on the data sheet.
    We are not sure if we should follow a specific protocol to get the
    staining work out fine.
    Thank you very much,

    Read More
    Answer

    Thank you for contacting us.

    I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    In order to better understand the problem, I would like to send you a questionnaire with some additional questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. Any images are highly appreciated.

    Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.

    I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

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