Western blot analysis was performed on membrane extracts (30 µglysate) of HeLa (Lane 1), and MCF-7 (Lane 2). The blots were probed with Rabbit Anti-SAP97 Polyclonal Antibody (Product # PA1-044, 2 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 110 kDa band corresponding to SAP97 was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues E(16) E Y R S K L S Q T E D R Q L R S S(33) of human SAP97.|
|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 *|
|Western Blot (WB)||2 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.
PA1-044 detects synapse-associated protein 97 (SAP97) from rat and human tissues. This antibody does not detect other synapse-associated protein family members.
PA1-044 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~140 kDa protein representing SAP97 from rat brain homogenate.
The PA1-044 immunogen is a synthetic peptide corresponding to residues E(16) E Y R S K L S Q T E D R Q L R S S(33) of human SAP97. This sequence is completely conserved in rat and mouse. PA1-044 immunizing peptide (Cat. # PEP-022) is available for use in neutralization and control experiments.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
SAP97 blocks the RXR ER retention signal of NMDA receptor subunit GluN1-3 through its SH3 domain.
PA1-044 was used in immunocytochemistry to explore how the different domains of SAP97 regulate its localization.
|Hong X,Avetisyan M,Ronilo M,Standley S||Biochimica et biophysica acta (1853:489)||2015|
Effect of rasagiline on the molecular composition of the excitatory postsynaptic density.
PA1-044 was used in western blot to investigate the influence of rasagiline on the ionotropic glutamate receptor composition of excitatory postsynaptic density
|Gardoni F,Zianni E,Eramo A,Canonico PL,Di Luca M||European journal of pharmacology (670:458)||2011|
Characteristics of the turnover of uncoupling protein 3 by the ubiquitin proteasome system in isolated mitochondria.
PA1-044 was used in western blot to investigate the role of uncoupling protein 3 in the ubiquitin proteasome system of mitochondria
|Mookerjee SA,Brand MD||Biochimica et biophysica acta (1807:1474)||2011|
Distinct levels of dopamine denervation differentially alter striatal synaptic plasticity and NMDA receptor subunit composition.
PA1-044 was used in western blot to investigate the effect of dopamine depletion on synaptic function and plasticity
|Paillé V,Picconi B,Bagetta V,Ghiglieri V,Sgobio C,Di Filippo M,Viscomi MT,Giampà C,Fusco FR,Gardoni F,Bernardi G,Greengard P,Di Luca M,Calabresi P||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:14182)||2010|