Immunofluorescent analysis of SAP97 (green) showing staining in the cytoplasm and membrane of NIH-3T3 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a SAP97 polyclonal antibody (Product # PA1-741) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues V(115) L P S E R I S P Q V P N E V L G P E(133) of rat SAP97.|
|Purification||Ammonium sulfate precipitation|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:1,000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:500-1:5000|
PA1-741 detects Synapse-Associated Protein 97 (SAP97) from human, rat and mouse tissues. No cross-reactivity to other proteins, including synapse-associated protein family members, has been observed.
PA1-741 has been successfully used in Western blot, immunofluorescence, immunocytochemistry, immunoprecipitation, and immunohistochemistry procedures. By Western blot, this antibody detects an ~140 kDa protein representing SAP97 from rat brain extract. Immunohistochemical staining of SAP97 in rat hippocampus with PA1-741 results in intense neuronal cell body and dendrite staining.
The PA1-741 immunizing peptide corresponds to amino acids 115-133 from rat SAP97. This sequence is 84% and 74% conserved in mouse and human SAP97, respectively.
Synapse-Associated Protein 97 (SAP97/human homologue of Drosophila discs-large tumor supressor or HDLG) is one of a family of plasma membrane-associated proteins found in synaptic junctions. This is a novel, presynaptic membrane protein homologous to SAP90 and the Drosophila discs-large tumor supressor protein. SAP97 has three ~90 amino acid repeats called PDZ domains, a single interior SH3 domain, and a COOH-terminal guanylate kinase homology (GuK) domain that is enzymatically inactive. It is hypothesized that PDZ-domain interactions play a role in receptor and channel clustering which contributes to neuronal plasticity. SAP97 is believed to participate in the clustering of certain proteins, including N-methyl-D-aspartate (NMDA) receptors and Shaker-type potassium channels at the synaptic membrane. NMDA receptors and shaker-type potassium channels both share C-terminal sequence homology consisting of a threonine/serine-X-valine-COOH (T/SXV) motif. Other neuronal proteins that share this motif (beta-1 adrenergic receptor, some serotonin receptors, some potassium channel subunits, and additional potassium channel subunits) may interact with SAP97 by binding to its PDZ domains. Neuronal nitric oxide synthase (nNOS), which lacks the T/SXV motif but which has its own PDZ domain, has been shown to associate with SAP97 in vitro through a pseudo-homotypic PDZ-PDZ interaction.
Structure-function analysis of SAP97, a modular scaffolding protein that drives dendrite growth.
PA1-741 was used in western blot to demonstrate that the dendrite branching promoting action of full length SAP97 is dependent on the PDZ3 domain
|Zhang L,Hsu FC,Mojsilovic-Petrovic J,Jablonski AM,Zhai J,Coulter DA,Kalb RG||Molecular and cellular neurosciences (65:31)||2015|
Postsynaptic density scaffold SAP102 regulates cortical synapse development through EphB and PAK signaling pathway.
PA1-741 was used in western blot to examine the roles of SAP102 in cortical synapse development
|Murata Y,Constantine-Paton M||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:5040)||2013|
Increased [³H]D-aspartate release and changes in glutamate receptor expression in the hippocampus of the mnd mouse.
PA1-741 was used in western blot to study the role of Cln8 gene in hippocampal excitabilit and the glutamatergic transmission system
|Bigini P,Milanese M,Gardoni F,Longhi A,Bonifacino T,Barbera S,Fumagalli E,Di Luca M,Mennini T,Bonanno G||Journal of neuroscience research (90:1148)||2012|
Cognitive impairment in humanized APP×PS1 mice is linked to Aβ(1-42) and NOX activation.
PA1-741 was used in western blot to investigate the relationship of cognitive impairment with NADPH oxidase activation and A beta deposition in a mouse Alzheimer disease model
|Bruce-Keller AJ,Gupta S,Knight AG,Beckett TL,McMullen JM,Davis PR,Murphy MP,Van Eldik LJ,St Clair D,Keller JN||Neurobiology of disease (44:317)||2011|
A specific requirement of Arc/Arg3.1 for visual experience-induced homeostatic synaptic plasticity in mouse primary visual cortex.
PA1-741 was used in western blot to investigate the importance of Arc/Arg3.1 in visual experience-induced synaptic plasticity in mouse primary visual cortex
|Gao M,Sossa K,Song L,Errington L,Cummings L,Hwang H,Kuhl D,Worley P,Lee HK||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:7168)||2010|
Differential recruitment of Kv1.4 and Kv4.2 to lipid rafts by PSD-95.
PA1-741 was used in western blot to study the role of PSD-95 in Kv channel recuitment and activity.
|Wong W,Schlichter LC||The Journal of biological chemistry (279:444)||2004|
Affinity purification of PSD-95-containing postsynaptic complexes.
PA1-741 was used in western blot to develop an affinity-based strategy for the purification of Triton-derived PSD fraction
|Vinade L,Chang M,Schlief ML,Petersen JD,Reese TS,Tao-Cheng JH,Dosemeci A||Journal of neurochemistry (87:1255)||2003|
Sequential delivery of synaptic GluA1- and GluA4-containing AMPA receptors (AMPARs) by SAP97 anchored protein complexes in classical conditioning.
PA1-741 was used in immunohistochemistry to study the role of SAP97 in the sequential delivery of synaptic GluA1 and GluA4 AMPAR subunits in classical conditioning
|Zheng Z,Keifer J||The Journal of biological chemistry (289:10540)||2014|
Pals1 is a major regulator of the epithelial-like polarization and the extension of the myelin sheath in peripheral nerves.
PA1-741 was used in immunohistochemistry to study the role of Pals1 in cellular polarization and the structure of myelin sheath in peripheral nerves
|Ozçelik M,Cotter L,Jacob C,Pereira JA,Relvas JB,Suter U,Tricaud N||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:4120)||2010|
Differential neuronal and glial expression of GluR1 AMPA receptor subunit and the scaffolding proteins SAP97 and 4.1N during rat cerebellar development.
PA1-741 was used in immunohistochemistry to study the expression of GluR1 AMPA receptor subunit and the scaffolding proteins SAP97 and 4.1N during rat cerebellar development
|Douyard J,Shen L,Huganir RL,Rubio ME||The Journal of comparative neurology (502:141)||2007|
A multiprotein trafficking complex composed of SAP97, CASK, Veli, and Mint1 is associated with inward rectifier Kir2 potassium channels.
PA1-741 was used in immunoprecipitation to demonstrate the CASK function in trafficking and plasma membrane localization of Kir2 channels.
|Leonoudakis D,Conti LR,Radeke CM,McGuire LM,Vandenberg CA||The Journal of biological chemistry (279:19051)||2004|
discs large homolog 1; discs large homolog 1, scribble cell polarity complex component; discs, large homolog 1; discs, large homolog 1 (Drosophila) transcript variant 6-v1; discs, large homolog 1 (Drosophila) transcript variant 6-v2; disks large homolog 1; Drosophila discs-large tumor suppressor homologue (synapse associated protein); embryo-dlg/synapse-associated protein 97; HDLG; Human Homologue of Drosophila Discs-Large Tumor Supressor; presynaptic protein SAP97; SAP-97; Synapse Associated Protein 97; synapse-associated protein 97
B130052P05Rik; dJ1061C18.1.1; DLG1; DLGH1; E-dlg/SAP97; hdlg; mKIAA4187; SAP-97; SAP97