- PA5-78733 - Invitrogen Antibodies ADO antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ADO in rat testis extract (lane 1), mouse kidney extract (lane 2) and HeLa whole cell lysate (lane 3). Sample was incubated with ADO polyclonal antibody (Product # PA5-78733) at a dilution of 0.5 µg/mL. Signal development was performed using a chemiluminescence (ECL) kit.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ADO in, Lane 1: rat brain tissue lysates, Lane 2: rat testicular tissue lysates, Lane 3: rat spleen tissue lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: human SW620 whole cell lysates, Lane 6: mouse spleen tissue lysates, Lane 7: mouse heart tissue lysates, Lane 8: mouse HEPA1-6 whole cell lysates, Lane 9: mouse SP2/0 whole cell lysates, . 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 50 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. The membrane was blocked with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with ADO Polyclonal Antibody (Product # PA5-78733) 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 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for ADO at approximately 28,30KD. The expected band size for ADO is at 30KD.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of ADO using anti-ADO antibody (Product # PA5-78733) . ADO was detected in a section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-ADO antibody (Product # PA5-78733) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow Cytometry of ADO in A549 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with ADO Polyclonal Antibody (Product # PA5-78733) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow Cytometry of ADO in PC-3 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with ADO Polyclonal Antibody (Product # PA5-78733) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry of ADO in U251 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with ADO Polyclonal Antibody (Product # PA5-78733) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry of ADO in HeLa cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with ADO Polyclonal Antibody (Product # PA5-78733) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

PA5-78733