Immunofluorescence analysis of SynGAP was performed using 70% confluent log phase PC-12 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SynGAP Rabbit Polyclonal Antibody (PA1046) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||GST fusion protein of a recombinant fragment, residues 947-1167, of rat SynGAP.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-046 detects synaptic Ras-GTPase activating protein (SynGAP) from mouse and rat tissues.
The PA1-046 has been successfully used in Western blot and immunocytochemistry procedures. By Western blot, this antibody detects an ~135 kDa protein representing SynGAP from mouse brain tissue homogenate. Immunocytochemical staining of SynGAP in rat hippocampal neurons with PA1-046 yields a pattern consistent with vesicular staining.
PA1-046 immunogen is residues 947-1167 of rat SynGAP.
Synaptic Ras-GTPase activating protein (SynGAP) is a PDZ-domain interacting protein expressed almost exclusively in the brain. SynGAP has been shown to be a major component of postsynaptic density and associates with post synaptic density 95 kDa (PSD-95), a synaptic scaffold protein, and synaptic N-methyl-D-aspartate (NMDA) receptors. SynGAP Ras-GTPase activity has also been shown to be inhibited by phosphorylation by CaM kinase II, another abundant protein located in the postsynaptic density. It has been proposed that SynGAP may function as a negative regulator of MAP Kinase activation in the absence of active NMDA receptors.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A model for regulation by SynGAP-¿1 of binding of synaptic proteins to PDZ-domain 'Slots' in the postsynaptic density.
PA1-046 was used in western blot demonstrate that synGAP-alpha1 regulates postsynaptic density composition by restricting binding to the PDZ domains of PSD-95
|Walkup WG,Mastro TL,Schenker LT,Vielmetter J,Hu R,Iancu A,Reghunathan M,Bannon BD,Kennedy MB||eLife (5:null)||2016|
The calcium sensor Copine-6 regulates spine structural plasticity and learning and memory.
PA1-046 was used in western blot to determine regulation of spine structural plasticity and memory and learning by the calcium sensor Copine-6
|Reinhard JR,Kriz A,Galic M,Angliker N,Rajalu M,Vogt KE,Ruegg MA||Nature communications (7:null)||2016|
GluN2B-Containing NMDA Receptors Regulate AMPA Receptor Traffic through Anchoring of the Synaptic Proteasome.
PA1-046 was used in immunocytochemistry and western blot to assess the role of the GluN2B subunit during development and in synaptic plasticity
|Ferreira JS,Schmidt J,Rio P,Águas R,Rooyakkers A,Li KW,Smit AB,Craig AM,Carvalho AL||The Journal of neuroscience : the official journal of the Society for Neuroscience (35:8462)||2015|
Impaired synaptic clustering of postsynaptic density proteins and altered signal transmission in hippocampal neurons, and disrupted learning behavior in PDZ1 and PDZ2 ligand binding-deficient PSD-95 knockin mice.
PA1-046 was used in western blot to use knock-in mice expressing PSD-95 lacking PDZ1/PDZ2 ligand binding to study the role of PSD-95 in the synaptic clustering of postsynaptic density proteins, synaptic transmission and memory
|Nagura H,Ishikawa Y,Kobayashi K,Takao K,Tanaka T,Nishikawa K,Tamura H,Shiosaka S,Suzuki H,Miyakawa T,Fujiyoshi Y,Doi T||Molecular brain (5:null)||2012|
Deletion of densin-180 results in abnormal behaviors associated with mental illness and reduces mGluR5 and DISC1 in the postsynaptic density fraction.
PA1-046 was used in western blot to investigate the role of densin-180 in the organization and dynamics of postsynaptic signaling complexes
|Carlisle HJ,Luong TN,Medina-Marino A,Schenker L,Khorosheva E,Indersmitten T,Gunapala KM,Steele AD,O'Dell TJ,Patterson PH,Kennedy MB||The Journal of neuroscience : the official journal of the Society for Neuroscience (31:16194)||2011|
In vivo composition of NMDA receptor signaling complexes differs between membrane subdomains and is modulated by PSD-95 and PSD-93.
PA1-046 was used in western blot to investigate the composition of NMDAR-PSD-95 complexes in lipid rafts
|Delint-Ramirez I,Fernández E,Bayés A,Kicsi E,Komiyama NH,Grant SG||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:8162)||2010|
Regulation of the neuron-specific Ras GTPase-activating protein, synGAP, by Ca2+/calmodulin-dependent protein kinase II.
PA1-046 was used in western blot to investigate the role of calcium/calmodulin-dependent protein kinase II in regulating synGAP.
|Oh JS,Manzerra P,Kennedy MB||The Journal of biological chemistry (279:17980)||2004|
A synaptic Ras-GTPase activating protein (p135 SynGAP) inhibited by CaM kinase II.
PA1-046 was used in western blot to characterize a synaptic Ras-GTPase activating protein in the brain
|Chen HJ,Rojas-Soto M,Oguni A,Kennedy MB||Neuron (20:895)||1998|
Phosphorylation of synaptic GTPase-activating protein (synGAP) by Ca2+/calmodulin-dependent protein kinase II (CaMKII) and cyclin-dependent kinase 5 (CDK5) alters the ratio of its GAP activity toward Ras and Rap GTPases.
PA1-046 was used in immunoprecipitation and western blot to study the effect of phosphorylation on synGAP activity
|Walkup WG,Washburn L,Sweredoski MJ,Carlisle HJ,Graham RL,Hess S,Kennedy MB||The Journal of biological chemistry (290:4908)||2015|
Cadherin-8 expression, synaptic localization, and molecular control of neuronal form in prefrontal corticostriatal circuits.
PA1-046 was used in immunohistochemistry to examine the progression of functional corticostriatal interactions
|Friedman LG,Riemslagh FW,Sullivan JM,Mesias R,Williams FM,Huntley GW,Benson DL||The Journal of comparative neurology (523:75)||2015|
Trans-synaptic EphB2-ephrin-B3 interaction regulates excitatory synapse density by inhibition of postsynaptic MAPK signaling.
PA1-046 was used in immunocytochemistry to investigate the mechanism of excitatory synapse density modulation by EphB2-ephrin-B3 association
|McClelland AC,Hruska M,Coenen AJ,Henkemeyer M,Dalva MB||Proceedings of the National Academy of Sciences of the United States of America (107:8830)||2010|
Age-related accumulation of Reelin in amyloid-like deposits.
PA1-046 was used in immunocytochemistry to examine the Reelin accumulation of Reelin in amyloid-like deposits during ageing
|Knuesel I,Nyffeler M,Mormède C,Muhia M,Meyer U,Pietropaolo S,Yee BK,Pryce CR,LaFerla FM,Marighetto A,Feldon J||Neurobiology of aging (30:697)||2009|
Synapse composition and organization following chronic activity blockade in cultured hippocampal neurons.
PA1-046 was used in immunocytochemistry to study the effect of activity on synapse composition and organization in central nerve system
|Harms KJ,Craig AM||The Journal of comparative neurology (490:72)||2005|
Gephyrin is critical for glycine receptor clustering but not for the formation of functional GABAergic synapses in hippocampal neurons.
PA1-046 was used in immunocytochemistry to study the GABAergic synaptic phenotype of hippocampal neurons from gephyrin-/- mice.
|Lévi S,Logan SM,Tovar KR,Craig AM||The Journal of neuroscience : the official journal of the Society for Neuroscience (24:207)||2004|
Dystroglycan is selectively associated with inhibitory GABAergic synapses but is dispensable for their differentiation.
PA1-046 was used in immunocytochemistry to investigate the functions of dystrophin glycoprotein complex in the differentiation of synapses
|Lévi S,Grady RM,Henry MD,Campbell KP,Sanes JR,Craig AM||The Journal of neuroscience : the official journal of the Society for Neuroscience (22:4274)||2002|