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

    21-30 of 39 Abreviews or Q&A

    Answer

    Thank you for your call today.

    I've issued credit note # *** for the order ***, and the original purchase price will be refunded back to your credit card pending approval of our accounting department. If you have any questions about this refund, please contact our accounting department at mailto:us.credits@abcam.com.

    As we discussed, please let me know once you have decided which antibodies you would like as replacements for ab32359 and the two orders of ab89170, and I will send those to you promptly. If you have any questions or would like assistance selecting antibodies for replacements, I will be happy to help you further.

    I look forward to hearing from you.

    Have a great day!

    Read More

    Answer

    Thank you for contacting us.

    Your credit note ID is 202-XXXX.

    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Question
    Answer

    Thank you for contacting us.

    The purchase order number is XXX.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    Thank you for contacting us on Friday in regards to Anti-S6K antibody [E175] (ab32359).
    I am very sorry for the change in specificity of ab32359, I understand your concern and the effect this will have on the interpretation of the results that you have previously collected. We try our very best to provide good quality reliable products but due to the variable nature of antibodies this is not always possible. When a problem is identified we perform a full investigation and inform the customers which have purchased the antibody as soon as we can.
    I am currently seeking further explanation from the source of this antibody in order to understand why the antibody is no longer deemed specific. I understand that the current problem is believed to affect at least the products purchased within the last year but may not be limited to these. I will let you know as soon as I have any further information relating to this.
    As discussed over the phone, I have also looked into the alternative antibodies that we have against the phospho T389 S6K. Currently we have rabbit polyclonal ab2571 and ab19380. Both of which have been raised against a phospho T398 peptide and have both been used in western blotting and validated with human samples.
    As can be seen from the datasheet of ab2571, the antibody has been used with AEE788 treated rat samples. This antibody has also been tested by using HeLa cells tested with and without l-phosphatase to test the specificity. I am currently seeking additional data for both ab2571 and ab19380 in how these antibodies have been validated.
    I can offer you either of these antibodies to try as an alternative to ab32359. Unfortunately we have neither of these antibodies in stock so it is likely to take up to a week for ab2571 to be delivered whilst ab19380 may take up to two weeks.
    I am very sorry for the inconvenience this is causing you and I will try to help you in whatever way I can to get your experiments up and running again.

    Read More

    Answer

    Thank you for contacting us.

    The PO number is XXX.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question
    Answer

    Thank you for contacting us.

    The PO number is XXX.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question
    Answer

    Thank you for contacting us.

    The PO number for this order is XXX.

    We have realized that this non specificity problem happened with batches produced in 2011. The batches before that period are absolutely fine, as of PO number XXX which was bought in 2006.

    Please ignore my previous email, this change should not have affected the end user experiment so I am closing this case. I apologize for the inconvenience this may have caused.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    Thank you for sending some further details on the protocol used for immunostaining and some images.

    The protocol seems to be absolutely fine but unfortunately there is no signal (for S6K) on the pictures you have kindly sent to me. I understand that you have used the recommended dilution (1/50) and ABC signal enhancing detection (biotinylated polyclonal swine anti-rabbit secondary antibody).

    The antibody may have been damaged during shipment or storage.

    I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    Since you have a working alternative primary antibody, you may not need a replacement vial. However, I could certainly arrange it for you, alternatively I could arrange a credit note which you can use for any future order.

    Please do let me know how you wish to proceed with your enquiry. I look forward to hearing from you soon.

    Read More

    Question
    Answer

    Thank you for contacting us.

    We recommend reduced and denaturing conditions in western blot.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question
    Answer

    Thank you very much for contacting us.
    I can confirm that I have sent an email to my colleague in lab and asked for more scientific proof of this change. I will get back to you soon, once I get a reply from them.
    Many thanks for having patience!

    Read More

    21-30 of 39 Abreviews or Q&A

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