P2Y12/P2RY12 Antibody (1C2A9) - BSA Free

Western Blot: P2Y12/P2RY12 Antibody (1C2A9) [NBP2-61749] - Analysis using P2RY12 mAb against HEK293 (1) and P2RY12 (AA: extra mix)-hIgGFc transfected HEK293 (2) cell lysate.

Flow Cytometry: P2Y12/P2RY12 Antibody (1C2A9) [NBP2-61749] - Analysis of Hela cells using P2RY12 mouse mAb (green) and negative control (red).

Western Blot: P2Y12/P2RY12 Antibody (1C2A9) [NBP2-61749] - Analysis using P2RY12 mAb against human P2RY12 (AA: extra mix) recombinant protein. (Expected MW is 9.4 kDa)

Western Blot: P2Y12/P2RY12 Antibody (1C2A9) [NBP2-61749] - Analysis using P2RY12 mouse mAb against PC-3 (1) and C6 (2) cell lysate.

ELISA: P2Y12/P2RY12 Antibody (1C2A9) [NBP2-61749] - Black line: Control Antigen (100 ng);Purple line: Antigen (10ng); Blue line: Antigen (50 ng); Red line:Antigen (100 ng)

P2Y12 regulates EGFR activation, SLUG and ZEB1 expression and enhances PDAC cells viability. (A) Immunoblots showing the expression of P2Y12, p-EGFR Y1068, EGFR, SLUG and ZEB1 in AsPC-1 and BxPC-3 after P2Y12-specific siRNA treatment. Cancer cells were seeded and treated with P2Y12 siRNA or a negative control siRNA, as described in the Methods and Materials section. (B) The columns represent the fold change of protein levels (p-EGFR Y1068, SLUG and ZEB1) relative to the negative control siRNA-treated cells, normalised to 1 (n ≥ 3). Statistical analysis was performed using one sample t-test (GraphPad Prism 8) comparing the mean of siP2Y12 treatment with the normalised value 1. (C) Cell viability of AsPC-1 and BxPC-3 cells following knockdown of P2Y12 compared with the negative control siRNA-treated cells (n ≥ 3), calculated using one sample t-test (Graphpad Prism 8) and the normalised control group mean of 100%. Data are presented as mean +/- SEM. *** p < 0.001, ** p < 0.002, * p < 0.033. siNeg: siRNA negative control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31968611), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The P2Y12 receptor, activated by ADP, triggers AKT activation in PDAC cells. (A) Immunoblots show the expression of P2Y12 and EGFR in PDAC cells (AsPC-1, BxPC-3, MiaPaCa-2, CFPAC-1 and PANC-1) and the normal pancreatic duct cells h-TERT-HPNE. Platelets were used as a positive control for P2Y12. Cells were seeded at 3 × 105 cells/well in a 6-well plate for 24 h, then washed, lysed and the proteins were collected and quantified. (B,C) Relative P2Y12 and EGFR expression in five PDAC cell lines and h-TERT-HPNE cells. The expression level was quantified and normalised to the loading control, alpha -actinin, with automated software Image Lab (version 5.1, BioRad, Hercules CA, USA) and represented as columns using GraphPad Prism 8 (GraphPad Software, Inc, CA, USA). (D) The P2Y12 inhibitor, ticagrelor, reduced ADP-induced AKT activation in AsPC-1 and BxPC-3. Briefly, cancer cells were seeded in a 12-well plate and after 24 h, cells were starved for 6 h, then treated with ticagrelor (5 uM) combined with ADP (100 uM) and the cells were further incubated for 30 min in serum-free media. The figure shows a representative blot from three independent experiments. (E) Extracellular ADP release from AsPC-1, BxPC-3, MiaPaCa-2 and h-TERT-HPNE. ADP was analysed in 250 uL of PBS previously incubated with cells for 15 min as described in the Methods and Materials section. The columns represent the mean of relative fluorescence units (RFU) from three independent experiments. (F) Western blot analysis of phospho-EGFR Y1068 (p-EGFR Y1068), phospho-AKT S473 (p-AKT S473) and phospho-ERK 1/2 (p-ERK1/2) expression in lysates derived from AsPC-1, BxPC-3 and h-TERT-HPNE treated with ADP (10, 50 and 100 uM) for 30 min. The figure shows a representative blot from three independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31968611), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu
• Applications: WB, ELISA, Flow
• Clone: 1C2A9
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

P2Y12/P2RY12 Antibody (1C2A9) - BSA Free Summary

• Immunogen: Purified recombinant fragment of human P2Y12/P2RY12 (AA: extra mix) expressed in E. Coli.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: P2RY12
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100-1:2000
  • Flow Cytometry 1:10-1:1000
  • Western Blot 1:100-1:2000
• Theoretical MW: 39.4 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for P2Y12/P2RY12 Antibody (1C2A9) - BSA Free

• ADPG-R
• Gi-coupled ADP receptor HORK3
• HORK3
• HORK3P2Y12 platelet ADP receptor
• P2RY12
• P2T(AC)
• P2Y purinoceptor 12
• P2Y(AC)
• P2Y(ADP)
• P2Y(cyc)
• P2Y12
• P2Y12ADP-glucose receptor
• purinergic receptor P2RY12
• purinergic receptor P2Y, G-protein coupled, 12
• putative G-protein coupled receptor
• SP1999
• SP1999G-protein coupled receptor SP1999

Background

The product of this gene belongs to the family of G-protein coupled receptors. This family has several receptor subtypes with different pharmacological selectivity, which overlaps in some cases, for various adenosine and uridine nucleotides. This receptor is involved in platelet aggregation, and is a potential target for the treatment of thromboembolisms and other clotting disorders. Mutations in this gene are implicated in bleeding disorder, platelet type 8 (BDPLT8). Alternative splicing results in multiple transcript variants of this gene.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for P2Y12/P2RY12 Antibody (NBP2-61749)(1)

Publication using NBP2-61749

• Elaskalani O, Domenichini A, Abdol Razak NB et al. Antiplatelet Drug Ticagrelor Enhances Chemotherapeutic Efficacy by Targeting the Novel P2Y12-AKT Pathway in Pancreatic Cancer Cells Cancers (Basel) 2020-01-20 [PMID: 31968611]

Bioinformatics

• Gene Symbol: P2RY12