711546

antibody from Invitrogen Antibodies

Targeting: P2RY12 HORK3, P2Y12, SP1999

Antibody data

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Validation data

P2RY12 protein structure - 711546 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of P2Y12 was achieved by transfecting PC3 cells with P2Y12 specific siRNA (Silencer® select Product # s34983 and Product # s34985). Western blot analysis (Fig a) was performed using TEB lysates from the P2Y12 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-P2Y12 Recombinant Rabbit Polyclonal Antibody (Product # 711546, 1-2 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Reduction of signal upon siRNA mediated knock down confirms that antibody is specific to P2Y12.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on Tissue extracts (30 µg lysate) of Mouse brain (Lane 1), Rat brain (Lane 2) and membrane extracts of U-87 MG (Lane 3), MCF-7 (Lane 4), MDA-MB-231 (Lane 5), PC-3 (Lane 6), IMR32 (Lane 7), T98G (Lane 8) and C8-D1A (Lane 9). The blots were probed with Anti-P2Y12 Recombinant Rabbit Polyclonal Antibody (Product # 711546, 1 µg/mL). A 60 kDa band corresponding to P2Y12 was observed across the cell lines and tissues tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).