Anti-IL-1RA antibody [EPR26853-58]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling IL-1RA with ab303490 at 1/5000 (0.09 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : No staining on human kidney. The section was incubated with ab303490 for 30 mins 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling IL-1RA with ab303490 at 1/100 (4.51 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear and cytoplasmic staining in A431 cell line.Negative control : MCF7 (PMID : 14581361)Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / A431 (human epidermoid carcinoma epithelial cell, Right) cells labelling IL-1RA with ab303490 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : MCF7(PMID : 14581361).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Immunohistochemical analysis of paraffin-embedded Human esophagus tissue labeling IL-1RA with ab303490 at 1/5000 (0.09 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human esophagus. The section was incubated with ab303490 for 30 mins 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling IL-1RA with ab303490 at 1/5000 (0.09 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human skin. The section was incubated with ab303490 for 30 mins 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.  Negative control : MCF7 (PMID : 14581361).

All lanes:

Western blot - Anti-IL-1RA antibody [EPR26853-58] (ab303490) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate (PMID:14581361) at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Observed band size: 20 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-IL-1RA antibody [EPR26853-58] (AB303490)

Blocking and dilution buffer and concentration : 5% NFDM/TBST.  Negative control : human kidney (human protein atlas database) In Western blot, ab303490 was shown to bind specifically to IL-1RA. A band was observed at 20kDa in wild-type A431 cell lysate with no signal observed at this size in IL-1RA knockout cell line (knockout cell lysate ab277366).

All lanes:

Western blot - Anti-IL-1RA antibody [EPR26853-58] (ab303490) at 1/1000 dilution

Lane 1:

Wild-type A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

IL-1RA knockout A431 whole cell lysate at 20 µg

Lane 3:

Human skin tissue lysate at 20 µg

Lane 4:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20 kDa

false

Exposure time: 59s