- PA5-95585 - Invitrogen Antibodies TNFRSF1A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of TNFR1 in Lane 1: human HeLa whole cell lysate, Lane 2: human K562 whole cell lysate, Lane 3: human Caco-2 whole cell lysate, Lane 4: rat liver tissue lysates, Lane 5: mouse small intestine tissue lysates, Lane 6: mouse liver tissue lysates, Lane 7: mouse HEPA1-6 whole cell lysate. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with TNFR1 polyclonal antibody (Product # PA5-95585) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of TNF Receptor I in, Lane 1: human Hela whole cell lysate, Lane 2: human K562 whole cell lysate, Lane 3: human Caco-2 whole cell lysate, Lane 4: rat liver tissue lysates, Lane 5: mouse small intestine tissue lysates, Lane 6: mouse liver tissue lysates, Lane 7: mouse HEPA1-6 whole cell lysate. 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 TNFR1 Polyclonal Antibody (Product # PA5-95585) 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 TNF Receptor I at approximately 50KD. The expected band size for TNF Receptor I is at 50KD.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of TNFR1 in paraffin-embedded mouse kidney tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with TNFR1 polyclonal antibody (Product # PA5-95585) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of TNFR1 in paraffin-embedded mouse liver tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with TNFR1 polyclonal antibody (Product # PA5-95585) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of TNFR1 in paraffin-embedded mouse lung tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with TNFR1 polyclonal antibody (Product # PA5-95585) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of TNFR1 in U937 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with TNFR1 polyclonal antibody (Product # PA5-95585) at a dilution of 1 µg (per 1x10^6 cells), followed by 488 conjugated goat anti-rabbit IgG (30 min at 20°C) using a 5-10 µg (per 1x10^6 cells) dilution.

PA5-95585