PA5-34822

antibody from Invitrogen Antibodies

Targeting: ROBO1 DUTT1, FLJ21882, SAX3

Antibody data

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Validation data

ROBO1 protein structure - PA5-34822 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ROBO1 using A) 30 µg HeLa whole cell lysate and B) 30 µg A375 whole cell lysate. Samples were loaded onto a 5% SDS PAGE gel and probed with a ROBO1 polyclonal antibody (Product # PA5-34822) at a dilution of 1:10000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of ROBO1 was achieved by transfecting HeLa with ROBO1 specific siRNAs (Silencer® select Product # s12092, s12093). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the ROBO1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ROBO1 Polyclonal Antibody (Product # PA5-34822, 1:1000 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 ROBO1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-ROBO1 Polyclonal Antibody (Product # PA5-34822) and a 200kDa band corresponding to ROBO1 was observed along with uncharacterized band (*) across cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), HUVEC (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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 (1:1000 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 Pico PLUS Chemiluminescent Substrate (Product # 34580).