Anti-IL-6R antibody [EPR24322-143]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-6R antibody [EPR24322-143] (AB271042)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling IL-6R with ab271042 at 1/2000 dilution (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cyoplasmics staining on human tonsil. The section was incubated with ab271042 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-6R antibody [EPR24322-143] (AB271042)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labelling IL-6R with ab271042 at 1/2000 dilution (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cyoplasmics staining on human placenta (PMID : 25658637). The section was incubated with ab271042 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IL-6R antibody [EPR24322-143] (AB271042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U266B1 cells labelling IL-6R with ab271042 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in U266B1 cells. Negative control : K-562 (PMID : 8300137) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-6R antibody [EPR24322-143] (AB271042)

Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labelling IL-6R with ab271042 at 1/2000 dilution (0.255 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cyoplasmics staining on human thyroid carcinoma. The section was incubated with ab271042 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

• IP

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Immunoprecipitation - Anti-IL-6R antibody [EPR24322-143] (AB271042)

IL-6R was immunoprecipitated from 0.35 mg U266B1 (human multiple myeloma B lymphocyte) whole cell lysate 10 ug with ab271042 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab271042 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : U266B1 (human multiple myeloma B lymphocyte) whole cell lysate 10 ug

Lane 2 : ab271042 IP in U266B1 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab271042 in U266B1 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5.5 seconds

All lanes:

Immunoprecipitation - Anti-IL-6R antibody [EPR24322-143] (ab271042)

Predicted band size: 52 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-IL-6R antibody [EPR24322-143] (AB271042)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The observed MW is consistent with what has been described in the literature (PMID : 28276471, 21282511).

Negative control : K-562 (PMID : 8300137)

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 37 seconds

All lanes:

Western blot - Anti-IL-6R antibody [EPR24322-143] (ab271042) at 1/1000 dilution

Lane 1:

U266B1 (human multiple myeloma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 70-80 kDa

false