- MA5-44916 - Invitrogen Antibodies UBE4A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Ube4A in different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. Samples were incubated in Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:500 in 5% BSA at room temperature for 2 hours followed by Goat Anti-Rabbit IgG - HRP secondary antibody at a dilution of 1:5,000 for 1 hour at room temperature. Positive control: Lane 1: HepG2 cell lysate; Lane 2: 293T cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Ube4A in Rat brain tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. Samples were incubated in Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:500 in 5% BSA at room temperature for 2 hours followed by Goat Anti-Rabbit IgG - HRP secondary antibody at a dilution of 1:5,000 for 1 hour at room temperature.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of Ube4A in PANC-1 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. Samples were incubated in Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:50 for 1 hour at room temperature, washed with PBS followed by Alexa Fluor®488 Goat anti-Rabbit IgG secondary antibody at a dilution of 1:1,000. The nuclear counter stain is DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of Ube4A in SK-OV-3 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. Samples were incubated in Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:50 for 1 hour at room temperature, washed with PBS followed by Alexa Fluor®488 Goat anti-Rabbit IgG secondary antibody at a dilution of 1:1,000. The nuclear counter stain is DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of Ube4A in SW620 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. Samples were incubated in Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:50 for 1 hour at room temperature, washed with PBS followed by Alexa Fluor®488 Goat anti-Rabbit IgG secondary antibody at a dilution of 1:1,000. The nuclear counter stain is DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Ube4A in paraffin-embedded human small intestine tissue. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1:50 for 30 minutes 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of Ube4A in HepG2 cells. The cells were fixed, permeabilized and stained with Ube4A Monoclonal antibody (Product # MA5-44916) using a dilution of 1 µg/mL (red) at 4℃ for 1 hour followed by Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG secondary antibody at a dilution of 1:1,000 for 30 minutes at 4℃. Rabbit IgG Isotype Control (green). Unlabeled sample was used as a control (cells without incubation with primary antibody; black).

MA5-44916