Anti-IL-11RA antibody [EPR5446]

10 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-11RA antibody [EPR5446] (ab125015)

  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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP) 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.

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  Immunocytochemistry/ Immunofluorescence - Anti-IL-11RA antibody [EPR5446] (ab125015)

  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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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.

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

  The expression pattern is consistent with what has been described in the literature (PMID: 26876177).
  

  Blocking Buffer and concentration: 5% NFDM/TBST

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

  Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 2: WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

  Lane 3: M1 (mouse myeloid leukemia myeloblast) whole cell lysate at 20 µg

  Lane 4: Mouse spleen tissue lysate at 20 µg

  Lane 5: Mouse testis tissue lysate at 20 µg

  Lane 6: Mouse thymus tissue lysate at 20 µg

  Lane 7: Mouse lymph node tissue lysate at 20 µg

  Secondary

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

  Predicted band size: 45 kDa

  Observed band size: 42 kDa, 45 kDa

  Exposure time: 3min

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

  All lanes: Western blot - Anti-IL-11RA antibody [EPR5446] (ab125015) at 1/10000 dilution

  Lane 1: A673 (Human muscle Ewing's Sarcoma ) whole cell lysates at 20 µg

  Lane 2: PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 3: HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates at 20 µg

  Lane 4: K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 20 µg

  Secondary

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

  Predicted band size: 45 kDa

  Observed band size: 45 kDa

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  Flow Cytometry (Intracellular) - Anti-IL-11RA antibody [EPR5446] (ab125015)

  Intracellular 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, Goat Anti-Rabbit IgG H&L (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).

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

  ELISA analysis of Human IL-11RA recombinant protein at 1000 ng/mL with ab125015. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

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

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

  Lane 1: 293T lysate at 10 µg

  Lane 2: Saos2 lysate at 10 µg

  Lane 3: A673 lysate at 10 µg

  Lane 4: PC3 lysate at 10 µg

  Lane 5: Hut78 lysate at 10 µg

  Lane 6: K562 lysate at 10 µg

  Lane 7: TF1 lysate at 10 µg

  Secondary

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

  Predicted band size: 45 kDa

  Observed band size: 45 kDa

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  Flow Cytometry (Intracellular) - Anti-IL-11RA antibody [EPR5446] (ab125015)

  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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-11RA antibody [EPR5446] (ab125015)

  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.

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  OI-RD Scanning - Anti-IL-11RA antibody [EPR5446] (ab125015)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.