Overview

  • Product name
    Anti-S6K1 antibody [E175]
    See all S6K1 primary antibodies
  • Description
    Rabbit monoclonal [E175] to S6K1
  • Host species
    Rabbit
  • Specificity
    ab32359 may detect both phosphorylated and non-phosphorylated forms of p70 S6 Kinase.
  • Tested applications
    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide within Human S6K1. The exact sequence is proprietary.

  • Positive control
    • Jurkat cell lysate; human breast carcinoma
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32359 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 1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).
IHC-P 1/50.
IP 1/60.
  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Acts to integrate nutrient and growth factor signals in regulation of protein synthesis, cell proliferation, cell growth, cell cycle progression and cell survival. Downstream effector of the mTOR signaling pathway. Phosphorylates specifically ribosomal protein S6 in response to insulin or several classes of mitogens. During translation initiation, the inactive form associatess with the eIF-3 complex under conditions of nutrient depletion. Mitogenic stimulation leads to phosphorylation and dissociation from the eIF-3 complex and the free activated form can phosphorylate other translational targets including EIF4B. Promotes protein synthesis by phosphorylating PDCD4 at 'Ser-67' and targeting it for degradation. Phosphorylates RICTOR leading to regulation of mammalian target of rapamycin complex 2 (mTORC2) signaling; probably phosphorylates RICTOR at 'Thr-1135'. Phosphorylates IRS1 at multiple serine residues coupled with insulin resistance; probably phosphorylates IRS1 at 'Ser-270'. Required for TNF-alpha induced IRS-1 degradation. Phosphorylates EEF2K in response to IGF1 and inhibits EEF2K activity. Phosphorylates BAD at 'Ser-99' in response to IGF1 leading to BAD inactivation and inhibition of BAD-induced apoptosis. Phosphorylates mitochondrial RMP leading to dissociation of a RMP:PPP1CC complex; probably phosphorylates RMP at 'Ser-99'. The free mitochondrial PPP1CC can dephosphorylate RPS6KB1 at Thr-412 which is proposed to be a negative feed back mechanism for the RPS6KB1 antiapoptotic function. Phosphorylates GSK3B at 'Ser-9' under conditions leading to loss of the TSC1-TSC2 complex. Phosphorylates POLDIP3.
    • Tissue specificity
      Widely expressed.
    • Sequence similarities
      Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.
      Contains 1 AGC-kinase C-terminal domain.
      Contains 1 protein kinase domain.
    • Domain
      The autoinhibitory domain is believed to block phosphorylation within the AGC-kinase C-terminal domain and the activation loop.
      The TOS (TOR signaling) motif is essential for activation by mTORC1.
    • Post-translational
      modifications
      Phosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism.
    • Cellular localization
      Cytoplasm; Nucleus. Cytoplasm and Cell junction > synapse > synaptosome. Mitochondrion outer membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 70 kDa ribosomal protein S6 kinase 1 antibody
      • KS6B1_HUMAN antibody
      • p70 alpha antibody
      • P70 beta 1 antibody
      • p70 ribosomal S6 kinase alpha antibody
      • p70 ribosomal S6 kinase beta 1 antibody
      • p70 S6 kinase alpha antibody
      • P70 S6 Kinase antibody
      • p70 S6 kinase, alpha 1 antibody
      • p70 S6 kinase, alpha 2 antibody
      • p70 S6K antibody
      • p70 S6K-alpha antibody
      • p70 S6KA antibody
      • p70(S6K) alpha antibody
      • p70(S6K)-alpha antibody
      • p70-alpha antibody
      • p70-S6K 1 antibody
      • p70-S6K antibody
      • P70S6K antibody
      • P70S6K1 antibody
      • p70S6Kb antibody
      • PS6K antibody
      • Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody
      • Ribosomal protein S6 kinase beta 1 antibody
      • Ribosomal protein S6 kinase beta-1 antibody
      • Ribosomal protein S6 kinase I antibody
      • RPS6KB1 antibody
      • S6K antibody
      • S6K-beta-1 antibody
      • S6K1 antibody
      • Serine/threonine kinase 14 alpha antibody
      • Serine/threonine-protein kinase 14A antibody
      • STK14A antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: S6K1 knockout HAP1 cell lysate (20 µg)
      Lane 3: MCF7 cell lysate (20 µg)
      Lane 4: HEK293 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab32359 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab32359 was shown to recognize S6K1 when S6K1 knockout samples were used, along with additional cross-reactive bands. Wild-type and S6K1 knockout samples were subjected to SDS-PAGE. ab32359 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Paraffin-embedded human breast carcinoma
      ab32359 at 1/50 dilution
    • Anti-S6K1 antibody [E175] (ab32359) + 293T cell lysate

      Predicted band size: 59 kDa

    References

    This product has been referenced in:
    • Noh JH  et al. Loss of RNA-binding protein GRSF1 activates mTOR to elicit a proinflammatory transcriptional program. Nucleic Acids Res 47:2472-2486 (2019). Read more (PubMed: 30753671) »
    • Cheng Z  et al. Rab1A promotes proliferation and migration abilities via regulation of the HER2/AKT-independent mTOR/S6K1 pathway in colorectal cancer. Oncol Rep 41:2717-2728 (2019). Read more (PubMed: 30896866) »
    See all 31 Publications for this product

    Customer reviews and Q&As

    31-39 of 39 Abreviews or Q&A

    Answer

    Thank you for your enquiry regardingab32359 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

    Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

    Read More

    Question
    Answer

    Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

    Unserem Wissen nach wurde ab2571 bisher noch nicht in IHC getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zurzeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab2571 abdecken, und gegen eine erneute Bestellung eines weiteren primären Antikörpers von uns verrechnet werden.

    Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

    1.) Bestätigen Sie mir bitte, dass Sie ab2571 inIHC testen möchten.

    2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

    3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

    4.) Testen Sie den Antikörper in IHC.

    5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
    Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
    https://www.abcam.com/abreviews

    6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderenprimären Antikörperzu dem gleichen Preis wie ab2571 bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer von ab2571 bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

    Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

    Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
    https://www.abcam.com/collaborationdiscount

    Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

    Read More
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Liver)
    Loading amount
    60 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 19 2012

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Fruit fly (Drosophila melanogaster) Tissue sections (Eye imaginal disc)
    Specification
    Eye imaginal disc
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - PBS 0.1% triton

    Abcam user community

    Verified customer

    Submitted Oct 13 2010

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (kidney)
    Specification
    kidney
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Oct 05 2007

    Answer

    Thank you for your enquiry. I have the exact testing conditions for use with this antibody and I will add this information to the online datasheet as soon as possible. The blocking and secondary was performed with milk, and the primary was diluted in BSA. The antibody originates outside of Abcam and was tested outside of Abcam. It is not a protocol that we usually follow in the lab at Abcam, but this is how the antibody was tested so I recommend that you follow this protocol. Please let me know if your result is successful follow this protocol or if you have further questions. Antibody Characterization through Western Immunoblotting 1. Block nitrocellulose for 1 hour at room temperature or overnight at 4 °C using 5% NFDM. 2. Incubate nitrocellulose with appropriate dilutions of primary antibody in 5% BSA overnight at 4 °C or for 2 hours at room temperature 3. Wash nitrocellulose with three 5-min washes using TBST. 4. Incubate nitrocellulose with 1:1000 (1:500 if stronger signal is desired) dilutions of goat anti-rabbit HRP antibody in 5% NFDM in TBST overnight at 4°C or room temperature for 1 hour. (Overnight at 4°C recommended for best results). 5. Wash nitrocellulose in 3 washes of TBST, then rinse in TBS prior to addition of chemiluminescence reagents. 6. Remove excess chemiluminescence reagent and sandwich nitrocellulose blot in any type of transparent plastic wrap (saran wrap, transparent sheet protector, etc.). 7. Acquire image using darkroom development techniques. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Answer

    Thank you for your enquiry. Based on the antibody originator's testing and the phospho site on the immunogen, this antibody should be specific to only p70 S6 kinase. In WB, this antibody showed only one band, specific for the p70 S6 kinase. However, they have not done any extensive testing to see if this antibody can also recognize p85 S6 kinase. Please note that other p70 S6 kinase antibodies have been shown to give both nuclear and cytosplasmic staining in certain cases. Also, p70 S6 kinase has been detected in the nucleus in some samples and conditions: Kim SJ, Kahn CR. Insulin stimulates p70 S6 kinase in the nucleus of cells. Biochem Biophys Res Commun. 1997;234:681-685. [PubMed] If you have any questions or comments, please feel free to contact me any time.

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    Answer

    Thank you for your enquiry. The antibody works well in IHC-P as evidenced by the image on the datasheet, but has not tested positive in ICC yet. In my experience it is strange for an antibody to work in IHC-P, but not ICC although this is possible and happens on rare occasions. One of the reasons why some of our antibodies may be negative by ICC but positive by IHC is that we usually try to use the same cell line that was used by western and IP. IF we do not get it to work (trying a different set of dilutions) we claim the antibody to be negative for that application. Another reason could be that there might be a better cell line to choose from for ICC, but we may not have access to that cell line, hence why we try to use what was positive by WB or IP. If you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abpoints, which can be redeemed on a number of rewards (a further 100 Abpoints will be offered for an image). Our Abpromise guarantee does not apply in this case. We cannot guarantee the antibody to work because in our hands it did not work in ICC. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question
    Answer

    Thank you for your enquiry and patience. This antibody has not been tested to determine whether it cross-reacts with the p85 S6 kinase or not. The originator of the antibody is going to re-test the antibody and we will let you know of the results. Please contact us again if you have any additional questions.

    Read More

    31-39 of 39 Abreviews or Q&A

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