Recombinant
RabMAb

Recombinant Anti-IL-11RA antibody [EPR5446] (ab125015)

Overview

  • Product name

    Anti-IL-11RA antibody [EPR5446]
    See all IL-11RA primary antibodies
  • Description

    Rabbit monoclonal [EPR5446] to IL-11RA
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human IL-11RA aa 350-450. The exact sequence is proprietary.

  • Positive control

    • WB: 293T, Saos2, A673, PC3, HuT78, K562, and TF1 cell lysates; IHC-P: Human kidney tissue; FC: K562 cells; ICC/IF: K562 cells;
  • General notes

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

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab125015 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 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified use in 1/500 - 1/1000

See IHC antigen retrieval protocols.

Flow Cyt 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use in 1/100 - 1/1000

ICC/IF 1/100.

For unpurified use in 1/500 - 1/1000

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Receptor for interleukin-11. The receptor systems for IL6, LIF, OSM, CNTF, IL11 and CT1 can utilize IL6ST for initiating signal transmission. The IL11/IL11RA/IL6ST complex may be involved in the control of proliferation and/or differentiation of skeletogenic progenitor or other mesenchymal cells.
    • Tissue specificity

      Expressed in a number of cell lines, including the myelogenous leukemia cell line K562, the megakaryocytic leukemia cell line Mo7E, the erythroleukemia cell line TF1, and the osteosarcoma cell lines, MG-63 and Saos-2. Also expressed in normal and malignant prostate epithelial cell lines. Expression levels are increased in prostate carcinoma.
    • Sequence similarities

      Belongs to the type I cytokine receptor family. Type 3 subfamily.
      Contains 2 fibronectin type-III domains.
      Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    • Cellular localization

      Membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • CRSDA antibody
      • hCG_2011440 antibody
      • I11RA_HUMAN antibody
      • IL-11 receptor subunit alpha antibody
      • IL-11R subunit alpha antibody
      • IL-11R-alpha antibody
      • IL-11RA antibody
      • IL11RA antibody
      • Interleukin 11 receptor alpha antibody
      • Interleukin 11 receptor alpha chain antibody
      • Interleukin-11 receptor subunit alpha antibody
      • MGC2146 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling IL-11RA with purified ab125015 at 1:250 dilution (3.2 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) ab97051
      was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling IL-11RA with Purified ab125015 at 1:100 dilution (8.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • All lanes : Anti-IL-11RA antibody [EPR5446] (ab125015) at 1/10000 dilution (Purified)

      Lane 1 : A673 (Human muscle Ewing's Sarcoma ) whole cell lysates
      Lane 2 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates
      Lane 3 : HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates
      Lane 4 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 45 kDa
      Observed band size: 45 kDa

    • Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling IL-11RA with Purified ab125015 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Overlay histogram showing K562 cells stained with unpurified ab125015 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab125015, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in K562 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • All lanes : Anti-IL-11RA antibody [EPR5446] (ab125015) at 1/1000 dilution (Unpurified)

      Lane 1 : 293T lysate
      Lane 2 : Saos2 lysate
      Lane 3 : A673 lysate
      Lane 4 : PC3 lysate
      Lane 5 : Hut78 lysate
      Lane 6 : K562 lysate
      Lane 7 : TF1 lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 45 kDa
      Observed band size: 45 kDa

    • Unpurified ab125015 at 1/500 staining IL-11RA in paraffin embedded Human Kidney tissue by immunohistochemistry.

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    ab125015 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Answer

    Thank you for your reply.

    Indeed, while this product is tested and guaranteed to work in Flow Cytometry, as the epitope is cytoplasmic, cells will need to be permeablized.

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

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    Answer

    Thank you for contacting us.

    The exact immunogen information for this product is considered proprietary and as such I am unable to share the sequence with you. I can however let you know that the immunogen used to create this product did correspond to the cytoplasmic domain of human Interleukin-11 receptor subunit alpha.

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

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    Answer

    Thank you for contacting us.

    For the WB image containing various cell line lysates, 10 ug of protein was loaded.
    5% non fat milk in TBST was used for blocking for 1 hr, Primary antibodu was used at a dilution of 1/1000 in 5%BSA/TBST overnight at 4 degrees.
    Secondary conjugated to HRP was used and developed after addition of chemiluminescence agents.


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

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