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Western blot analysis of Human IL-18 on different lysates with Rabbit anti-Human IL-18 antibody (HA721767) at 1/1,000 dilution.
Lane 1: U-2 OS cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (20 µg/Lane)
Lane 4: HepG2 cell lysate (20 µg/Lane)
Predicted band size: 22 kDa
Observed band size: 20 kDa
Exposure time: 1 minute 45 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721767) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Human IL-18 antibody (HA721767) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721767) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Human IL-18 antibody (HA721767) at 1/10,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721767) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Human IL-18 antibody (HA721767) at 1/10,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721767) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Application: IF-Tissue
Species: Human
Site: spleen
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000
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Human IL-18 was immunoprecipitated from 0.2 mg A549 cell lysate with HA721767 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721767 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: A549 cell lysate (input)
Lane 2: Rabbit IgG instead of HA721767 in A549 cell lysate
Lane 3: HA721767 IP in A549 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 5 seconds; ECL: K1801
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