Recombinant Anti-IL-11RA antibody [EPR5446] - BSA and Azide free (ab240950)

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).

This data was developed using ab125015, the same antibody clone in a different buffer formulation.

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125015).

All lanes : 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
Lane 2 : WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate
Lane 3 : M1 (mouse myeloid leukemia myeloblast) whole cell lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Mouse testis tissue lysate
Lane 6 : Mouse thymus tissue lysate
Lane 7 : Mouse lymph node tissue lysate

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: 42,45 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

This data was developed using ab125015, the same antibody clone in a different buffer formulation.

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

Blocking Buffer and concentration: 5% NFDM/TBST

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125015).

This data was developed using ab125015, the same antibody clone in a different buffer formulation.

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.

Unpurified ab125015 at 1/500 staining IL11RA in paraffin embedded Human Kidney tissue by immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125015).

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

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125015).

Equilibrium disassociation constant (K_D)
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125015).