The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/500. Predicted molecular weight: 106 kDa.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/50 - 1/100.
Is unsuitable for IP.
Receptor for GABA. The activity of this receptor is mediated by G-proteins that inhibit adenylyl cyclase activity, stimulates phospholipase A2, activates potassium channels, inactivates voltage-dependent calcium-channels and modulates inositol phospholipids hydrolysis. Plays a critical role in the fine-tuning of inhibitory synaptic transmission. Pre-synaptic GABA-B-R inhibit neurotransmitter release by down-regulating high-voltage activated calcium channels, whereas postsynaptic GABA-B-R decrease neuronal excitability by activating a prominent inwardly rectifying potassium (Kir) conductance that underlies the late inhibitory postsynaptic potentials. Not only implicated in synaptic inhibition but also in hippocampal long-term potentiation, slow wave sleep, muscle relaxation and antinociception.
Highly expressed in brain, especially in cerebral cortex, thalamus, hippocampus, frontal, occipital and temporal lobe, occipital pole and cerebellum, followed by corpus callosum, caudate nucleus, spinal cord, amygdala and medulla. Weakly expressed in heart, testis and skeletal muscle.
Belongs to the G-protein coupled receptor 3 family. GABA-B receptor subfamily.
Alpha-helical parts of the C-terminal intracellular region mediate heterodimeric interaction with GABA-B receptor 1.
Cell membrane. Cell junction > synapse > postsynaptic cell membrane. Moreover coexpression of GABA-B-R1 and GABA-B-R2 appears to be a prerequisite for maturation and transport of GABA-B-R1 to the plasma membrane.
ab75838, at a 1/100 dilution, staining GABA B Receptor 2 in paraffin embedded human brain tissue by Immunohistochemistry.
Anti-GABA B Receptor 2 antibody [EP2411Y] (ab75838)
Western blot - GABA B Receptor 2 antibody [EP2411Y] (ab75838)
Anti-GABA B Receptor 2 antibody [EP2411Y] (ab75838) at 1/500 dilution + human cerebellum lysate at 10 µg
Secondary goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 106 kDa Observed band size: 106 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABA B Receptor 2 antibody [EP2411Y] (ab75838)This image is courtesy of an Abreview submitted by Estella Munoz.
Ab75838 staining GABA B receptor 2 in Rat Postnatal (P15) Cerebellum by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a 10mM citrate buffer, pH6. Samples were incubated with primary antibody (1/400 in PBS with 2% donkey serum) for 16 hours at 25°C. An Alexa Fluor® 488 conjugated Donkey anti-rabbit, polyclonal was used as the secondary antibody.
IHC and confocal microscopy for GABA B Receptor 2 (GABAB2) in the postnatal rat cerebellum. (A, C, E, G) Negative control by omission of the primary antibody at 10, 20, 40 and 60X. (B-B’’, D-D’’, F-F’’, H-H’’) Immunolabeling for GABAB2 (green) and/or the calcium-binding protein calbindin (red) at 10, 20, 40 and 60X. (I-K) Zooms of the insets shown in F-F’’. Specific signal for GABAB2 is mainly observed in the molecular layer (ML) and internal granular layer (IGL) of the cerebellar grey matter. Purkinje cells (PkL) are enriched in GABAB2 especially at the surface of the dendritic trees (white arrowheads) oriented towards the ML. Golgi II cells (yellow arrowhead) located in the IGL are also positive for GABAB2. EGL: external granular layer. n: calbindin-positive nucleus of a Purkinje cell. Orange arrowhead: GABAB2-immunoreactive granular cell in the IGL. WM: white matter. An Olympus FV 1000 confocal microscope was used and images were processed with Adobe Photoshop CS4. Figure generated by Juan Vilchez Aruani and Estela Muñoz, IHEM, UNCuyo, CONICET, Mendoza, Argentina.
Immunocytochemistry/ Immunofluorescence - Anti-GABA B Receptor 2 antibody [EP2411Y] (ab75838)Image from Cuny H et al., J Biol Chem. 2012 Jul 6;287(28):23948-57. Epub 2012 May 21.; Fig 2.; doi: 10.1074/jbc.M112.342998; July 6, 2012, The Journal of Biological Chemistry, 287, 23948-23957.
Immunofluorescence analysis of GABA B Receptor 2 using ab75838 at 1/400 dilution. Left panel = normal rat DRG neurons. Right panel = rat DRG neurons transfected with siRNAs to disrupt GABA B Receptor 2.
Flow Cytometry-Anti-GABA B Receptor 2 antibody [EP2411Y](ab75838)
Overlay histogram showing SH-SY5Y cells stained with ab75838 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75838, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Shen W et al. GABAB receptor attenuation of GABAA currents in neurons of the mammalian central nervous system. Physiol Rep5:N/A (2017).
Read more (PubMed: 28348006) »
Jiang Y et al. Opposing needling promotes behavior recovery and exerts neuroprotection via the cAMP/PKA/CREB signal transduction pathway in transient MCAO rats. Mol Med Rep13:2060-70 (2016).
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