Anti-IL1RA/Il1rn Antibody Picoband®

Western blot analysis of IL1RA/Il1rn using anti-IL1RA/Il1rn antibody (A00651-5).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: rat spleen tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat thymus tissue lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse spleen tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse RAW264.7 whole cell lysates,
Lane 9: mouse NIH/3T3 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-IL1RA/Il1rn antigen affinity purified polyclonal antibody (Catalog # A00651-5) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IL1RA/Il1rn at approximately 18KD. The expected band size for IL1RA/Il1rn is at 18KD.

Flow Cytometry analysis of HEPA1-6 cells using anti-IL1RA/Il1rn antibody (A00651-5).
Overlay histogram showing HEPA1-6 cells stained with A00651-5 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-IL1RA/Il1rn Antibody (A00651-5, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of NRK cells using anti-IL1RA/Il1rn antibody (A00651-5).
Overlay histogram showing NRK cells stained with A00651-5 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-IL1RA/Il1rn Antibody (A00651-5, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of IL1RA/Il1rn using anti-IL1RA/Il1rn antibody (A00651-5).
IL1RA/Il1rn was detected in a paraffin-embedded section of mouse lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IL1RA/Il1rn Antibody (A00651-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of IL1RA/Il1rn using anti-IL1RA/Il1rn antibody (A00651-5).
IL1RA/Il1rn was detected in a paraffin-embedded section of rat lung tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IL1RA/Il1rn Antibody (A00651-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.