rabbit polyclonal anti-rac2 antibody (Santa Cruz Biotechnology)
Structured Review

Rabbit Polyclonal Anti Rac2 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti-rac2 antibody/product/Santa Cruz Biotechnology
Average 90 stars, based on 22 article reviews
Images
1) Product Images from "GPR55 regulates cannabinoid 2 receptor-mediated responses in human neutrophils"
Article Title: GPR55 regulates cannabinoid 2 receptor-mediated responses in human neutrophils
Journal: Cell Research
doi: 10.1038/cr.2011.60
Figure Legend Snippet: GPR55 activation suppresses the CB2R-mediated activation and translocation of Rac2. (A) Neutrophils were stimulated with agonists (1 μM) for 1 min at 37 °C. The active GTP-bound Rac2 was extracted from the lysates with PAK domain-gluthatione agarose beads. GTP-bound and total GTPase levels were visualized by western blotting using a rabbit anti-Rac2 antibody, β-actin served as a loading control. The ratio of GTP-bound vs total GTPase levels was assessed with ImageJ software (graph). Data are mean±SEM of three independent experiments. (*P< 0.05; ***P< 0.001). (B) Serum-starved dHL60 cells were stimulated with agonists (1 μM) for 1 min at 37 °C. The extraction of active GTP-bound Rac2 was performed as in panel (A). Data are mean±SEM of three independent experiments (**P< 0.01; ***P< 0.001). (C) Neutrophils were seeded on fibronectin-coated glass coverslips and treated with 0.01% DMSO (control; i) or ligands for 5 min at 37 °C. Fixed cells were incubated with rabbit anti-Rac2 antibody and stained with Alexa Fluor-488 goat anti-rabbit antibody (green), Texas-Red Phalloidin (red), and DAPI (blue). Control (i) and LPI (3 μM, ii) treated cells show a nuclear/perinuclear localization of Rac2 (arrows). Upon 2-AG stimulation (1 μM, iii), Rac2 distributed evenly in the cytosol (arrow) and partially colocalized with actin at the plasma membrane (yellow, arrowhead). (iv) Treatment with a combination of LPI (3 μM) and 2-AG (1 μM) showed a nuclear localization of Rac2 in polarized neutrophils. Cells were analyzed using an OLYMPUS fluorescence microscope equipped with a Hamamatsu ORCA CCD camera (original magnification: 60×). Scale bars: 10 μm. Representative images from 2-3 experiments are shown.
Techniques Used: Activation Assay, Translocation Assay, Western Blot, Software, Incubation, Staining, Fluorescence, Microscopy
Figure Legend Snippet: Crosstalk between GPR55 and CB2R and its consequent biological responses in human blood neutrophils. (right panel) Stimulation of CB2R and GPR55 by 2-AG and LPI, respectively, leads to a coordinated activation of RhoA, Rac1 and Cdc42 small GTPases. This leads to a distinct remodeling of cytoskeleton (polarization) compared to sole activation of each receptor and facilitates neutrophils migration towards the gradient of agonists. (left panel) The bacterial killing mechanisms, which are provoked by C5a or 2-AG, are mediated via the activation of Rac2 small GTPases. Active Rac2 will translocate and incorporate to the NADPH oxidase core complex in the phagocytic cup and catalyzes the formation of reactive oxygen species (ROS). On the other hand, it facilitates degranulation of neutrophils via translocation of azurophilic granules, containing myeloperoxidase (MPO). Stimulation of GPR55 by LPI, via a yet unknown mechanism, inhibits activation of Rac2, thereby limiting the ROS production and degranulation.
Techniques Used: Activation Assay, Migration, Translocation Assay
