IHC staining of purified anti-P2RY12 antibody (clone S16007D) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve‐All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was incubated with the primary antibody at 5 µg/mL overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 647 conjugated Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI. The image was captured with a 10X objective. Scale bar: 100 µm
IHC staining of purified anti-P2RY12 antibody (clone S16007D) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve‐All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was incubated with the primary antibody at 5 µg/mL overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 647 conjugated Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI. The image was captured with a 10X objective. Scale bar: 100 µm
IHC staining of purified anti-P2RY12 antibody (clone S16007D) on formalin-fixed paraffin-embedded mouse brain tissue. Following antigen retrieval using Retrieve‐All Antigen Unmasking System 3 (Cat. No. 927801), the tissue was incubated with the primary antibody at 5 µg/mL overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 647 conjugated Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI. The image was captured with a 40X objective. Scale bar: 50 µm
Single cell suspension of mouse brain tissue was stained with purified anti-P2RY12 antibody (clone S16007D, blue histogram) or rat IgG isotype control (red histogram), followed by anti-rat antibody conjugated to APC for detection.
IHC staining of purified anti-P2RY12 antibody (clone S16007D) on 4%-formaldehyde fixed mouse brain tissue. Following thick (100µm) vibratome sectioning, the slices were blocked for 1h at RT with 10%NGS and 1%BSA in PBST (pH=7.4 PBS + 0.1% Tween20), incubated with the primary antibody at 5 µg/ml overnight at 4°C, followed by incubation with 2.5 µg/ml of Alexa Fluor® 647 conjugated Goat anti-rat IgG (Cat. No. 405416) for 2h at RT. CX3CR1-GFP signal enabled validation of the microglial-specific anti-P2RY12 staining (pseudocolored red). The above images are maximal intensity projections from 40µm-thick z-stacks captured with a 60X objective on a confocal laser scanning microscope. The images are a courtesy of Dr. Alberto Sepulveda-Rodriguez and Dr. Stefano Vicini, Department of Pharmacology and Physiology, Interdisciplinary Program in Neuroscience, Georgetown University Medical Center.
Purinergic Receptor P2Y (P2RY12) is a receptor for ADP and ATP coupled to G-proteins that inhibit the adenylyl cyclase second messenger system. P2RY12 is not activated by UDP and UTP. Required for normal platelet aggregation and blood coagulation. P2RY12 is a highly selective marker for microglia that specifically distinguishes these cells from other myeloid cells.
Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5.0 - 10 µg/ml is suggested. For flow cytometric staining. a concentration range of 0.5 - 5.0 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes
View the video of our collaborator discussing the importance of highly specific monoclonal antibodies for P2RY12 in Microglia research.
Learn more about the validation testing we performed on clone S16007D by watching our Scientific Poster Video.
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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