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

    11-20 of 39 Abreviews or Q&A

    Answer

    Thank you for your telephone enquiry.

    I am sorry there have been some difficulties regarding the specificity of ab32359S6K antibody [E175]

    As discussed on the telephone, I would be pleased to provide you with an alternative free of charge replacement. For example wehave the following S6K phospho T389 antibodies tested for WB and human. I can recommend to review the online datasheets for further information, then let me know which antibody you would like to receive.

    ab2571Anti-S6K (phospho T389) antibody
    Rabbit polyclonal
    Tested in: WB
    Reacts: with Human, Mouse, Rat
    https://www.abcam.com/index.html?datasheet=2571 (or use the following: https://www.abcam.com/index.html?datasheet=2571).

    ab19380 Anti-S6K (phospho T389) antibody
    Rabbit polyclonal
    Tested in: WB
    Reacts with: Human
    https://www.abcam.com/index.html?datasheet=19380 (or use the following: https://www.abcam.com/index.html?datasheet=19380).

    Reviewing the details on our records,I can confirm that we sent our product information to customera who hadpurchased ab32359 asafter conducting number of tests, our source for this product concluded that this antibody detects both phosphorylated and non-phosphorylated forms of p70 S6 Kinase. I apologize for any inconvenience this has caused.

    I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question
    Answer

    Es tut mir leid, dass Ihnen dieser Antikörper so viele Probleme bereitet hat. Es ist in der Tat so, dass dieser Antikörper seine Phosphoprotein- Spezifität verloren hat. Wir haben daher eine Informationskampagne gestartet, in der wir unseren Kunden eine Gutschrift angeboten haben. Auch Frau XXXXX hat solch eine Email erhalten, und zwar am 23.03.12.

    "At Abcam we aim to let our customers know everything that we do about the products in our catalogue. All the data we possess are shown on our datasheets and these are updated daily with any new information we’ve received, whether this is from customers, suppliers or our own labs. When this new information is something that may affect the quality of a product, we also contact relevant customers as a priority, to share this information and offer our assistance where necessary.

    We have recently received information regarding ab32359; Rabbit monoclonal [E175] to S6K which you have previously purchased from us. They have informed us that there has been an error, and this antibody is not phospho specific in contrast to previously info on the datasheet. We realize this information may impact your data assessment, and we would like to apologize for the inconvenience.

    The quality of our antibodies is very important to us, as are the feedback and data we receive. It is regrettable that the information we had on this particular antibody was not of the high standard both we and our customers expect from our products. Should this information impact the assessment of your data and you wish to accept compensation, please contact me with your original order number, and I will be pleased to arrange an immediate refund or credit note.

    Should you have any further questions regarding this issue, please do not hesitate to contact me."

    Es tut mir leid, dass diese Information Sie nicht erreicht hat, aber wir stellen Ihnen natürlich gerne wie am Telefon besprochen einen Ersatzantikörper oder eine Gutschrift aus.

    Read More

    Answer

    Thank you for contacting us.

    I have received the following explanation from the supplying lab:

    After conducting a number of tests, wemade the conclusion that the antibody detects both phosphorylated and non-phosphorylated forms of p70 S6 Kinase.

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

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    Thank you for contacting us.

    I am still in touch with the lab to inquire about the details of the change in the antibody target for ab32359.

    As soon as I receive the requested informationI will let you know.

    Thank you for your patience and understanding.

    In the meantime, please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    I am sorry for the delay in getting back to you with more news regarding the S6K antibody ab32359.
    I now have some data which I can share with you which will hopefully reassure you of the quality of the antibody and enable you to reassess the results collected.
    As I have mentioned previously. The lot numbers GR23148-1 and GR40447-1 were from the same batch produced and were the first lots identified as having increased cross-reactivity with the non-phospho protein. The extent of this cross reactivity was assessed through performing a dot blot using the T389 phospho modified and unmodified peptide (Figure 1). As can clearly be seen from this comparison, whilst the antibody is able to detect both sequences, the affinity of the antibody is higher towards the phospho-modified peptide. The non-selectivity can also be observed with FBS and serum starved Jurkat cell lysates (Figure 2). This data is the reason why the specificity of the antibody has been altered on the datasheet of ab32359 and why we decided to issue a product recall. We are sorry that this problem was not identified earlier and the implications that this has had on your work.
    The data for the lots preceding this batch seem to be vastly more specific towards the T389 phospho S6K. Figure 3 illustrates the results obtained with lot G15283 (-1 and -2) and 926308 both of which you have used (Purchase orders GEM3821703, GEM3826221, GEM3830007 and GEM3808410). These batches compare well to the originally produced antibody (and the figure previously presented on the datasheet of ab32359), Figure 3 blot furthest to the right.
    We think the difference between the results observed with GR23148-1 and GR40447-1 and the earlier batches is that the antibody was originally produced by the hybridoma cell line (clone E175). The antibody produced by this cell line was then taken to be produced as a recombinant protein. The decision was taken to develop the antibody in this way as it can lead to improved yields and concentration of the antibody produced. However, it seems that in this case it has also affected the specificity of the antibody towards it s target. Another possibility would be a rare mutation of the clone, that might have affected the affinity of this clone to the non-phospho protein. We are still investigating into this matter. We are intending to re-isolate the original hybridoma in order to reproduce the specific antibody again.
    I hope this information goes some way towards reassuring you that whilst the lot GR23148-1 and GR40447-1 are regrettably not specific towards the T398 phospho S6K, the preceding lots showed good selectivity (comparable to the data presented previously on the datasheet of ab32359).
    I understand the impact this will have had on the analysis of your work and my manager has agreed to let me offer you two free antibodies of your choice from our catalogue. I hope this goes some way towards compensating for the situation.
    If you have any further questions please do not hesitate to contact me.

    Read More

    Answer

    Thank you for getting back to me.
    I have organised for the credit notes to be issued for orders xxxxxx. The credit note IDs are as follows: xxxxxxxx. The credit notes 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.
    The credit note IDs are 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 now have evidence from the lab that the lot numbers GR23148-1 and GR40447-1 (discussed in my previous email) were detecting both the non-phosphorylated S6K as well as the phosphorylated form. I am still waiting on further confirmation as to how far back this problem extends. I am sorry for the delay in getting this information to you and any inconvenience this is causing.
    As mentioned in my previous email, we only take liability for the cost of the faulty product itself but I can understand your frustration with the situation. I am currently discussing with my manager what kind of extra compensation we may able to offer you.
    I will get back to you as soon as I have any further information.

    Read More

    Answer

    ab32359 detects both forms of p70S6K,phospho and non-phospho.

    Read More

    Answer

    Thank you for providing me with the order details up to the 24th October 2010.
    As discussed over the phone, I understand your frustrations and the level of resources and work time which has been used with this antibody. I am very sorry for this situation and the problems it is causing you. As I mentioned, when we are aware that there may be a problem with one of our antibodies, we are proactive and try to notify our customers as soon as possible.
    We are still looking into the problem in more depth but with the limited information I currently have from the lab it seems that it is a problem affecting the lot number GR23148-1 and GR40447-1 and the lots produced following this batch. You received these lots on the 25/02/2011, 30/03/2011 and 29/07/2011. As soon as I have further information to share with you I will send it on. I am happy to refund you directly for the three lots which are definitely affected (purchase order numbers GEM3837151, GEM3843667 and GEM3864297). In order to do this, could you confirm if I can refund it to the account used to pay for each of these orders? Or would you prefer a credit note for each order?
    I will confirm as quickly as I can how the activity has been assessed with the preceding orders and if these lots were also affected. If this turns out to be the case I will arrange for a refund of these too.
    The Abpromise guarantee which covers all the products in our catalogue promises to replace or refund any product which does not perform as stated on the datasheet. More information on this can be found from the following link:
    www.abcam.com/abpromise
    Details of our terms and conditions can also be found from the following page link:
    https://www.abcam.com/index.html?pageconfig=terms
    This states that:
    “Abcam will notify its customers as soon as practically possible after it becomes aware of deficiencies in a product supplied. Any claim relating to products shall be limited to replacement or refund of original purchase price paid.”
    I’d like to reassure you that we are taking this seriously and I have raised this case with my manager.

    Read More

    Answer

    Thank you for contacting us.

    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.


    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

    11-20 of 39 Abreviews or Q&A

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