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☑ Relative expression (RE)
Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.
Lane 1: U-87 MG cell lysate
Lane 2: A549 cell lysate
Lane 3: SK-Br-3 cell lysate (low expression)
Lane 4: MDA-MB-231 cell lysate
Lane 5: Mouse brain tissue lysate
Lane 6: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 103 kDa
Observed band size: 130 kDa
Exposure time: 35 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 (ET1609-69) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/200 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 (ET1609-69) at 1/200 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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☑ Relative expression (RE)
Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/5,000 dilution.
Lane 1: A549 cell lysate
Lane 2: MDA-MB-231 cell lysate
Lane 3: SK-Br-3 cell lysate (low expression)
Lane 4: Mouse brain tissue lysate
Lane 5: Mouse heart tissue lysate
Lane 6: Rat brain tissue lysate
Lane 7: Rat heart tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 103 kDa
Observed band size: 130 kDa
Exposure time: Lane 1-3: 1 minute 21 seconds; Lane 4-7: 43 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 (ET1609-69) at 1/5,000 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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ICC staining of Neuropilin 1 in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Neuropilin 1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Neuropilin 1 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-69, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/50 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 (ET1609-69) at 1/50 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 mouse kidney tissue with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/200 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 (ET1609-69) at 1/200 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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☑ Knockdown (KD)
Western blot analysis of Neuropilin 1 on different lysates with Rabbit anti-Neuropilin 1 antibody (ET1609-69) at 1/5,000 dilution.
Lane 1: MDA-MB-231-si NT cell lysate
Lane 2: MDA-MB-231-si Neuropilin 1 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 103 kDa
Observed band size: 130 kDa
Exposure time: 1 minute 2 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 (ET1609-69) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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