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    s6k1-antibody-ab9366.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors Rb
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Anti-S6K1 antibody (ab9366)

  • Datasheet
Reviews (8)Q&A (9)References (25)

Product price, shipping and contact information

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Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Key features and details

  • Rabbit polyclonal to S6K1
  • Suitable for: WB
  • Reacts with: Mouse
  • Isotype: IgG

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Overview

  • Product name

    Anti-S6K1 antibody
    See all S6K1 primary antibodies
  • Description

    Rabbit polyclonal to S6K1
  • Host species

    Rabbit
  • Specificity

    Reacts strongly with 70 kd of p70 S6K, but only weakly with the 80 kDa S6K.
  • Tested Applications & Species

    Application Species
    WB
    Mouse
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human S6K1 (C terminal) conjugated to keyhole limpet haemocyanin.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 6.00
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Immunoaffinity chromatography with S6K C-terminal peptide on agarose.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Rb
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • S6K Family
    • Metabolism
    • Types of disease
    • Obesity

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human S6K1 protein (ab62291)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab9366 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
WB
Mouse
Application Abreviews Notes
WB (8)
Use at an assay dependent concentration. Predicted molecular weight: 59 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 59 kDa.

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.
  • Target information above from: UniProt accession P23443 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 72508 Mouse
    • SwissProt: Q8BSK8 Mouse
    • Unigene: 394280 Mouse
    • Unigene: 446624 Mouse
    • Unigene: 479484 Mouse
    • 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

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (25)

    Publishing research using ab9366? Please let us know so that we can cite the reference in this datasheet.

    ab9366 has been referenced in 25 publications.

    • Zha A  et al. The Evaluation of the Antioxidant and Intestinal Protective Effects of Baicalin-Copper in Deoxynivalenol-Challenged Piglets. Oxid Med Cell Longev 2020:5363546 (2020). PubMed: 32064026
    • Wang X  et al. TFEB-NF-?B inflammatory signaling axis: a novel therapeutic pathway of Dihydrotanshinone I in doxorubicin-induced cardiotoxicity. J Exp Clin Cancer Res 39:93 (2020). PubMed: 32448281
    • Lai WS & Ding YL GNG7 silencing promotes the proliferation and differentiation of placental cytotrophoblasts in preeclampsia rats through activation of the mTOR signaling pathway. Int J Mol Med 43:1939-1950 (2019). PubMed: 30864685
    • Yang Y  et al. Skullcapflavone I suppresses proliferation of human lung cancer cells via down-regulating microRNA-21. Exp Mol Pathol 110:104285 (2019). PubMed: 31301304
    • Ma YF  et al. Phosphorylation of AKT serine/threonine kinase and abundance of milk protein synthesis gene networks in mammary tissue in response to supply of methionine in periparturient Holstein cows. J Dairy Sci 102:4264-4274 (2019). PubMed: 30879806
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 17 Abreviews or Q&A

    Question

    Hi,

    Thank you for your email.

    The customer wants to purchase AB12864 and I have let them know about the testing discount.

    We would be interested in the testing discount.

    We will be testing AB12864 in mouse and sheep, for Western Blot. Sample is kidney lysate.

    Their details are;

    Could you please confirm if they qualify for this?

    Many thanks,

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 23 2012

    Answer

    Thank you for contacting us.

    Yes the customer would be eligible for a testing discount for ab12864 if they are willing to test it in mouse and/or sheep samples with western blotting. The sequence surrounding the Serine 235 and 236 of human RPS6 (SwissProt http://www.uniprot.org/uniprot/P62753), against which the antibody was raised, shares 100% homology with the mouse protein (SwissProt reference http://www.uniprot.org/uniprot/P62754) and we would therefore expect it to react. I was unable to find a sequence for the sheep protein but it seems that the sequence is highly conserved in mammals and I would therefore expect the antibody also to be able to detect the sheep protein.

    In order for me to issue a testing discount could you please provide me with the email address and institute that the customer is with.

    I look forward to receiving your reply.

    Read More

    Abcam Scientific Support

    Answered on Mar 23 2012

    Western blot abreview for Anti-S6K1 antibody

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (BRAIN)
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (4-20% gradient)
    Loading amount
    40 µg
    Specification
    BRAIN
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 27 2019

    Western blot abreview for Anti-S6K1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Bos taurus Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (4-15% Gradient)
    Loading amount
    20 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Julie Kim

    Verified customer

    Submitted May 05 2017

    Question

    Sehr geehrte Damen und Herren,
    ich arbeite mit dem Anti-S6K- Antikörper (ab9366)um S6K in der humanen
    Magenkrebszelllinie MKN45 nachzuweisen. Leider ist es mir nicht
    möglich, eine klare Bande bei ca.70kDa nachzuweisen. Stattdessen habe
    ich Banden bei 70,40 und 10kDa und einen sehr starken Hintergrund. Da
    ich dachte, dass der AK eventuell zu stark sein könnte, habe ich ihn
    bereits in , wie empfohlen, einer 1:250 Verdünnung, 1:500 und auch
    1:1000 Verdünnung benutzt. Leider sind die Ergebnisse nicht besser
    geworden.Haben Sie eventuell eine Idee, was ich noch verändern/besser
    machen könnte?Die Bilder meiner Westernblots habe ich Ihnen im Anhang
    beigefügt. Auch eine ausführliche Beschreibung meines Protokolls.
    Vielen Dank für Ihre Hilfe.
    Mit freundlichen Grüßen

    Read More

    Abcam community

    Verified customer

    Asked on Jul 09 2012

    Answer

    Vielen Dank für Ihre Anfrage und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen. Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten. Ich habe gesehen, dass Sie schon mit meiner Kollegin bezüglich des ab2571 in Kontakt waren. Haben Sie das Problem mit dem ab2571 gelöst? Wir würden uns sehr freuen zu hören, wie Sie vorangekommen sind.

    In Bezug auf den ab9366,möchte ich gernedie folgenden Punkte ansprechen und Vorschlägezur Veränderungdes Protokolls machen.

    1.) Können Sie bestätigen, dass der sekundäre Antikörper ansonsten gut funktioniert auf diesen Zelllysaten? Gibt der sekundäre Antikörper auch Hintergrundbanden?

    2.) Ich kann Ihnen empfehlen, die Zelllysate in einem RIPA Puffer herzustellen. Die Wahl des Puffers ist wichtig, da einige Puffer gewisse Proteine nicht genug gut in Lösung bringen können. RIPA ist ein starker und guter Puffer. In unserem Onlineprotokoll haben wir auch einen Absatz über die Wahl der Puffer: https://www.abcam.com/index.html?pageconfig=resource&rid=11379#A1

    3.) Ich kann auch empfehlen, Inhibitoren von Proteasen den Lysaten hinzufügen.

    4.) Ich kann auch empfehlen, die Proben zu reduzieren. Falls wir keine spezielle Notiz auf dem Datenblatt haben, wurde der Antikörper unter reduzierten Kondition getestet.

    Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Wie Sie wissen,garantieren wiralle Produkte so zu funktionieren wie auf dem Datenblatt beschrieben. Falls wir also das Problem nicht lösen können, werden wir Ihnen diesen Antikörperersetzen.

    Ich freue mich wieder von Ihnen zu hören.

    Read More

    Abcam Scientific Support

    Answered on Jul 09 2012

    Western blot abreview for Anti-S6K1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Liver)
    Loading amount
    30 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 02 2012

    Question

    Hi,

    For some reason they email address did not copy on my email!

    The customer’s details are:

    xxxxx
    Thanks!

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 26 2012

    Answer

    Thank you for providing those details.


    I can now confirm the discount code is xxxxx for which the expiration date is the 24th of July 2012. In order to activate the code, please submit an Abreview for ab12864 using mouse and/or sheep samples with western blotting, quoting the discount code in the "additional comments" section. More information on this offer can be found from the link:

    https://www.abcam.com/collaboratordiscount

    If you have any further questions, please do not hesitate to contact us again. I wish you all the best with your research.

    Read More

    Abcam Scientific Support

    Answered on Mar 26 2012

    Question

    Hi xxx,

    Thank you for your help, please see below the customer’s response;

    Thank you for following up our antibody problem with Abcam. It is disturbing to hear that we may have a faulty vial as we have spent a lot of our time trying to optimise this antibody. I would like to take up the offer of a replacement, but I am reluctant to try another S6K antibody as we have already tried three with little success (one to the phosphorylated form and two not ).

    Another antibody commonly used to probe this pathway is the target of S6K called ribosomal protein S6. I am hoping that antibodies to this protein will give us better results. Abcam has 21 antibodies to RPS6. It is difficult to ascertain which ones will be successful. They need to be in stock as we have a paper currently under consideration with a journal and this data will enable it to be accepted.

    We need two antibodies, that recognize the phosphorylated and the non-phosphorylated form, respectively. The antibody ab40820 or ab59261 may be suitable as the non-phosphorylated option, and ab65748 or ab12864 for the phosphorylated form.

    Can you please advise which are in stock and would be likely to give us a good result on tissue lysates in a Western blot.

    Hope to hear from you soon!

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 19 2012

    Answer

    Thank you for getting back to me.

    I understand your concerns over the antibody. I suggested that the antibody may be at fault not because we have had other complaints from customers but because of the optimisation having been performed by the customer and the highly non-specific blot obtained. However, if you have used a further two antibodies to this target with similar results it may be that the samples do not contain the protein as expected. It may be if they have been stored at -80 for a long time, or gone through any freeze thaw cyclesthat they have degraded with time. It may be worthwhile to test the antibodies used with a positive control such as 293T cell lysate or jurkat cell lysate if you have these to hand? If you do not I would be able to provide this for you.

    If you would prefer to take a replacement antibody, I can have this organised too. I have had a look at the antibodies suggested against RPS6. Both ab40820 and ab5961 seem like good choices against the unphosphorylated target. These antibodies have both been validated in western blotting and with mouse samples and would therefore both be covered by the Abpromise. From the antibodies chosen against the phosphorylated RPS6target, ab65748 and ab12864, I would suggest ab65748 would be the most appropriate to use as this has been validated with mouse samples which ab12864 has not (we can only say that we would expect it to work due to the sequence homology of the mouse protein with the immunogen used but we cannot guarantee it).

    For all of the antibodies suggested, nonehave been used to detect the sheep protein and I cannot therefore say whether they would besuitable for performing western blottingwith thesesamples.

    I hope this information has been of help. I look forward to hearing how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on Mar 19 2012

    Question

    Hi xxxxx,

    Please see below the response from the customer;

    The S6K antibodies recommended storing at 4C & have been stored there.

    3. The no primary did yield some bands & I will send the blot

    4. The secondary antibody was Sigma A0545 anti-rabbit peroxidase antibody.

    5. They were all fresh membranes & this was the only antibody used on them.

    I've attached the Western blot image. The marker we use is Invitrogen Magic marker. The the strips are the same, marker, sheep, mouse.

    Please let me know if you require any further information

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 14 2012

    Answer

    Thank you for taking time to complete our questionnaire and for providing me with that extra information. I am sorry that this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. I apologize for the inconvenience has caused your customer and am pleased to offer you a free of charge replacement, credit note, or refund in compensation. Unfortunately, we only have the same lot of ab9366 in stock at the moment. I would therefore suggest https://www.abcam.com/S6K-antibody-6B2-ab119252.htmlwhich is a mouse monoclonal or https://www.abcam.com/S6K-antibody-ab47504.htmlwhich is a rabbit polyclonal. Both have been validated in WB and with mouse samples.

    I would still be veryinterested in seeing the "no primary" control result, just to see if some improvements could be made in improving this non-specificity..

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Abcam Scientific Support

    Answered on Mar 14 2012

    Western blot abreview for Anti-S6K1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (Liver)
    Loading amount
    50 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 13 2012

    Western blot abreview for Anti-S6K1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Liver)
    Loading amount
    50 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 13 2012

    1-10 of 17 Abreviews or Q&A

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