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Signal Transduction Protein Phosphorylation Ser / Thr Kinases S6K Family
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

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  • Certificate of Compliance
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Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Flow Cytometry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
  • Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1982Y] to RSK4 - BSA and Azide free
  • Suitable for: ICC, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

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Recombinant human RSK4 protein (Active) (ab85296)

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Overview

  • Product name

    Anti-RSK4 antibody [EP1982Y] - BSA and Azide free
    See all RSK4 primary antibodies
  • Description

    Rabbit monoclonal [EP1982Y] to RSK4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC, IHC-P, Flow Cyt, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human RSK4 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q9UK32

  • General notes

    ab247441 is the carrier-free version of ab76117 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab247441 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1982Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • S6K Family

Associated products

  • Alternative Versions

    • Anti-RSK4 antibody [EP1982Y] (ab76117)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
  • Recombinant Protein

    • Recombinant human RSK4 protein (Active) (ab85296)

Applications

Our Abpromise guarantee covers the use of ab247441 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
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 90 kDa (predicted molecular weight: 84 kDa).

Target

  • Function

    Serine/threonine kinase that may play a role in mediating the growth-factor and stress induced activation of the transcription factor CREB.
  • 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 2 protein kinase domains.
  • Post-translational
    modifications

    Autophosphorylated on Ser-389, as part of the activation process.
  • Target information above from: UniProt accession Q9UK32 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 27330 Human
    • Omim: 300303 Human
    • SwissProt: Q9UK32 Human
    • Unigene: 234478 Human
    • Alternative names

      • 90 kDa ribosomal protein S6 kinase 6 antibody
      • KS6A6_HUMAN antibody
      • p90 RSK 6 antibody
      • p90-RSK 6 antibody
      • p90RSK6 antibody
      • pp90RSK4 antibody
      • Ribosomal protein S6 kinase 90kDa polypeptide 6 antibody
      • Ribosomal protein S6 kinase alpha 6 antibody
      • Ribosomal protein S6 kinase alpha-6 antibody
      • Ribosomal S6 kinase 4 antibody
      • rps6ka6 antibody
      • RSK 4 antibody
      • RSK-4 antibody
      • S6K-alpha-6 antibody
      see all

    Images

    • Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

      Lane 1: Wild-type HAP1 cell lysate (40 µg)

      Lane 2: RSK4 knockout HAP1 cell lysate (40 µg)

      Lane 3: MCF7 cell lysate (20 µg)

      Lane 4: HEK293 cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab76117 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab76117 was shown to recognize RSK4 when RSK4 knockout samples were used, along with additional cross-reactive bands. Wild-type and RSK4 knockout samples were subjected to SDS-PAGE. ab76117 and ab8245 (loading control to GAPDH) were diluted at 1/250 and 1/10,000 dilution respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      All lanes : Anti-RSK4 antibody [EP1982Y] (ab76117) at 1/1000 dilution (purified)

      Lane 1 : SH-SY5Y cell lysate
      Lane 2 : Caco-2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 84 kDa
      Observed band size: 90 kDa
      why is the actual band size different from the predicted?



      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

      Immunofluorescence staining of HEK293 cells with purified ab76117 at a working dilution of 1/1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab76117 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

      Immunohistochemical staining of paraffin embedded rat kidney with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

      Immunohistochemical staining of paraffin embedded mouse cerebral cortex with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      This data was developed using ab76117, the same antibody clone in a different buffer formulation.Immunohistochemical staining of paraffin embedded human kidney with purified ab76117 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Flow Cytometry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Flow Cytometry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      This data was developed using ab76117, the same antibody clone in a different buffer formulation.Overlay histogram showing Neuro-2a cells fixed in 2% PFA and stained with purified ab76117 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
    • Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Western blot - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Anti-RSK4 antibody [EP1982Y] (ab76117) at 1/500 dilution (unpurified) + SH-SY5Y cell lysate at 10 µg

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 84 kDa
      Observed band size: 90 kDa why is the actual band size different from the predicted?



      This data was developed using ab76117, the same antibody clone in a different buffer formulation.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      This data was developed using ab76117, the same antibody clone in a different buffer formulation.Unpurified ab76117, at 1/50 dilution, staining RSK4 in paraffin-embedded human brain glioma tissue. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
    • Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Immunocytochemistry - Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      This data was developed using ab76117, the same antibody clone in a different buffer formulation.Immunofluorescent staining of SH-Sy5y cells using unpurified ab76117 at 1/100 dilution.
    • Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)
      Anti-RSK4 antibody [EP1982Y] - BSA and Azide free (ab247441)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab247441? Please let us know so that we can cite the reference in this datasheet.

    ab247441 has not yet been referenced specifically in any publications.

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