Invitrogen

P2Y12 Recombinant Rabbit Monoclonal Antibody (4H5L19)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

2 Published Figures

5 References

View all (27) P2Y12 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

P2Y12 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of P2Y12 (red) in human iPSC-derived microglia. The cells were fixed with 4% paraformaldehyde for 15 min at room temperature, and then permeabilized and blocked with 0.25% Triton X-100 and 10% donkey serum in PBS for 20 min. Samples were then incubated with Anti-P2Y12 Recombinant Rabbit Monoclonal Antibody (red; Product # 702516) at a dilution of 1:500 in PBS containing 0.25% Triton X-100 and 10% donkey serum at 48304C overnight, followed by incubation with Donkey Anti-Rabbit Alexa Fluor 568 (Product # A10042) at a dilution of 1:1000 as well as DAPI (blue; 1:25000) in PBS containing 0.25% Triton X-100 and 10% donkey serum at room temperature for 1 hour. Images were taken at 20X magnification. (Scale bar: 50 µm) Data courtesy of Dr. Zhexing Wen at Emory University.

P2Y12 Antibody in Western Blot (WB) — Western blot analysis was performed on Tissue extracts (30 µg lysate) of Rat Brain (Lane 1), Mouse Brain (Lane 2) and membrane extracts of U-87 MG (Lane 3), MCF-7 (Lane 4), MDA-MB-231 (Lane 5), PC-12 (Lane 6), IMR32 (Lane 7), SK-N-AS (Lane 8) and C8-D1A (Lane 9). The blots were probed with Anti-P2Y12 Recombinant Rabbit Monoclonal Antibody (Product # 702516, 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, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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).

P2Y12 Antibody in Western Blot, Immunocytochemistry, Immunohistochemistry (WB, ICC/IF, IHC) — Figure 1 The P2Y12 receptor is expressed in mouse models for liver cirrhosis and hepatocellular carcinoma. ( A ) Immunohistochemical staining with P2Y12 antibodies of liver sections from healthy mice, mice with carbon tetrachloride (CCL 4) -induced liver cirrhosis and mice with diethylnitrosamine (DEN)-induced liver cancer. ( B ) Quantification of immunohistochemical staining with P2Y12 antibodies. ( C ) Co-staining of P2Y12 and macrophage marker F4/80 in liver slides from DEN-induced hepatocellular carcinoma (HCC). ( D ) Immunocytochemical staining of THP1-differentiated macrophages and Raw264.7 cells. ( E ) Western Blot image of P2Y12 protein expression and beta-actin protein in THP1-differentiated macrophages and Raw264.7 cells. ( F ) mRNA expression levels of P2yr12 in interferon gamma and lipopolysaccharide-stimulated anti-tumoral (M1) macrophages and interleukin (IL)-13 and IL-4-stimulated pro-tumoral (M2) macrophages ( G ) Quantification of protein expression in pro-tumoral (M2) and anti-tumoral (M1) macrophages. ( H ) Representative image of Western blot showing P2Y12 protein expression in pro-tumoral (M2) and anti-tumoral (M1) macrophages. ( I ) mRNA expression of P2yr12 in livers from healthy mice, mice with CCL4-induced liver cirrhosis and mice with DEN-induced liver cancer. * = p < 0.05; ** = p < 0.01.

P2Y12 Antibody in Immunohistochemistry (IHC) — Figure 2 Expression of P2Y12 is located in the stroma of patients with hepatocellular carcinoma. ( A ) Micrographs of hepatocellular carcinoma biopsies stained with P2Y12 antibodies, obtained from the Human Protein Atlas, marking tumor (T) and stomal (S) area [,]. ( B ) Detail of stroma and ( C ) tumor areas within the biopsies []. ( D ) Kaplan-Meier survival curve of HCC patients with high or low expression of P2Y12 obtained through the Human Protein Atlas []. ( E ) P2Y12 expression in different HCC stages, measured as number fragments per kilobase of exon per million reads (RPKM) from the Human Protein Atlas [].

P2Y12 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. PC-3 cells were transfected with P2Y12 siRNA and decrease in signal intensity was observed in Western blot application (Fig a) using Anti-P2Y12 Recombinant Rabbit Monoclonal Antibody (Product # 702516). Densitometric analysis of this Western blot is shown in histogram (Fig b). {KD}

Product Details

702516

Applications	Tested Dilution	Publications
Western Blot (WB)	1 µg/mL	View 2 publications 2 publications
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	Assay-dependent	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Recombinant Monoclonal
Type	Antibody
Clone	4H5L19
Immunogen	Peptides corresponding to human P2Y12 [1) aa323-aa342, 2) aa119-aa138] View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2689476

Product Specific Information

This antibody is predicted to react with Monkey, Cat, Dog

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

P2Y12 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 platelets aggregation, and is a potential target for the treatment of thromboembolisms and other clotting disorders. Two transcript variants encoding the same isoform have been identified for this gene. Expression of P2Y12 has been reported in platelets, spinal cord, and many brain regions. ESTs for P2Y12 have been isolated from B-cell/lung/testis, brain, embryo, prostate, eye, kidney carcinoma, and colon carcinoma libraries.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Crosstalk between astrocytes and microglia results in increased degradation of α-synuclein and amyloid-β aggregates.

Journal of neuroinflammation

Rostami J,Mothes T,Kolahdouzan M,Eriksson O,Moslem M,Bergström J,Ingelsson M,O'Callaghan P,Healy LM,Falk A,Erlandsson A

702516 was used in Immunocytochemistry to show that intracellular deposits of αSYN and Aβ are significantly reduced in co-cultures of astrocytes and microglia, compared to monocultures of either cell type.

Thu Jun 03 00:00:00 EDT 2021

Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype.

International journal of molecular sciences

Pavlović N,Kopsida M,Gerwins P,Heindryckx F

Published figure using P2Y12 recombinant monoclonal antibody (Product # 702516) in Immunocytochemistry

Sat Oct 31 00:00:00 EDT 2020

Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype.

International journal of molecular sciences

Pavlović N,Kopsida M,Gerwins P,Heindryckx F

Published figure using P2Y12 recombinant monoclonal antibody (Product # 702516) in Immunocytochemistry

Sat Oct 31 00:00:00 EDT 2020

Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype.

International journal of molecular sciences

Pavlović N,Kopsida M,Gerwins P,Heindryckx F

Published figure using P2Y12 recombinant monoclonal antibody (Product # 702516) in Immunocytochemistry

Sat Oct 31 00:00:00 EDT 2020

Elevated protein synthesis in microglia causes autism-like synaptic and behavioral aberrations.

Nature communications

Xu ZX,Kim GH,Tan JW,Riso AE,Sun Y,Xu EY,Liao GY,Xu H,Lee SH,Do NY,Lee CH,Clipperton-Allen AE,Kwon S,Page DT,Lee KJ,Xu B

702516 was used in Western Blotting to propose that functional perturbation of male microglia is an important cause for sex-biased autism spectrum disorders (ASD).

Tue Apr 14 00:00:00 EDT 2020

Bioinformatics

Protein Aliases: ADP-glucose receptor; ADPG-R; G-protein coupled receptor SP1999; Gi-coupled ADP receptor HORK3; P2T(AC); P2Y purinoceptor 12; P2Y(AC); P2Y(ADP); P2Y(cyc); P2Y12; P2Y12 platelet ADP receptor; P2YR12; purinergic receptor P2RY12; purinergic receptor P2Y, G-protein coupled, 12; putative G-protein coupled receptor; SP1999

Gene Aliases: 2900079B22Rik; 4921504D23Rik; ADPG-R; BDPLT8; HORK3; P2RY12; P2T(AC); P2Y(12)R; P2Y(AC); P2Y(ADP); P2Y(cyc); P2Y12; SP1999

UniProt ID: (Human) Q9H244, (Mouse) Q9CPV9, (Rat) Q9EPX4

Entrez Gene ID: (Human) 64805, (Mouse) 70839, (Rat) 64803

Molecular Function: G-protein coupled receptor