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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- 711546 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- P2Y12 Recombinant Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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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.
- 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).
