- PA5-95551 - Invitrogen Antibodies DDR2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DDR2 in Lane 1: rat kidney tissue lysates, Lane 2: mouse kidney tissue lysates. 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 DDR2 polyclonal antibody (Product # PA5-95551) 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 was performed using Anti-DDR2 Polyclonal Antibody (Product # PA5-95551) and a 130 kDa band corresponding to Ddr2 was observed in IMR-32, SH-SY5Y, HT-1080 and BJ, which are reported to be positive cell models for Ddr2 but not in MCF7 and SK-OV-3, which are negative cell models. Membrane enriched extracts (30 µg lysate) of IMR-32 (Lane 1), SH-SY5Y (Lane 2), HT-1080 (Lane 3), BJ (Lane 4), MCF7 (Lane 5) and SK-OV-3 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). An uncharacterized band (*) at ~70 kDa was observed in all cell models and other uncharacterized bands (*) were observed at ~100, 55 and 38 kDa in some cell models.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Ddr2 was achieved by transfecting BJ with Ddr2 specific siRNAs (Silencer® select Product # S9762, S9761). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the Ddr2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DDR2 Polyclonal Antibody (Product # PA5-95551, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Ddr2. An uncharacterized band (*) at ~100 kDa was observed in all lysates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of DDR2 in paraffin-embedded mouse small intestine 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 DDR2 polyclonal antibody (Product # PA5-95551) at a 2 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 DDR2 in paraffin-embedded rat small intestine 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 DDR2 polyclonal antibody (Product # PA5-95551) at a 2 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

PA5-95551