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12882
Neuronal Scaffold Proteins Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Neuronal Scaffold Proteins Antibody Sampler Kit #12882

Citations (2)
Simple Western™ analysis of lysates (0.1 mg/mL) from mouse brain using PSD95 (D27E11) XP® Rabbit mAb #3450. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
Western blot analysis of extracts from the indicated rat brain synaptic fractions using SHANK2 Antibody. Separation of the different synaptic fractions was confirmed using PSD95 (D27E11) XP® Rabbit mAb #3450 and Syntaxin 6 (C34B2) Rabbit mAb #2869. Equal loading of each fraction was assessed using β-Tubulin (9F3) Rabbit mAb #2128. Fractionation of the different synaptic compartments was carried out as described by Phillips, G.R. et al. (2001) Neuron 32, 63-77. PAZ, Pre-synaptic active zone.
Western blot analysis of extracts from various tissues and cell lines using Spinophilin (E1E7R) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from mouse and rat brain tissue using Homer1 Antibody.
Western blot analysis of extracts from mouse and rat brain tissue using SHANK2 Antibody.
Immunoprecipitation of spinophilin from mouse brain extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Spinophilin (E1E7R) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Spinophilin (E1E7R) Rabbit mAb.
Confocal immunofluorescent analysis of rat cerebellum and retina using PSD95 (D27E11) XP® Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using Spinophilin (E1E7R) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded MCF7 (left) and Huh7 (right) cell pellets using Spinophilin (E1E7R) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Spinophilin (E1E7R) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded rat brain using Spinophilin (E1E7R) Rabbit mAb.
Confococal immunofluorescent analysis of MCF7 (left), SH-SY5Y (center), or Huh7 (right) cells using Spinophilin (E1E7R) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Inquiry Info.# 12882

Product Description

The Neuronal Scaffold Proteins Antibody Sampler Kit provides an economical means of evaluating four major scaffolding proteins. The kit includes enough primary antibody to perform four western blot experiments.

Specificity / Sensitivity

PSD95 (D27E11) XP® Rabbit mAb detects endogenous levels of total PSD95 protein. SHANK2 Antibody recognizes endogenous levels of total SHANK2 protein. Homer1 Antibody detects endogenous levels of total Homer1 protein. This antibody may also detect isoform Homer1c. Spinophilin (E1E7R) Rabbit mAb recognizes endogenous levels of total spinophilin protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val537 of human SHANK2 protein or residues surrounding Glu130 of human Homer1 protein. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly99 of human PSD95 or residues surrounding Pro298 of human spinophilin protein.

Background

Scaffold proteins are composed of protein-interaction domains that tether multiple components of a signaling pathway to form signal transduction complexes. This organization of signaling molecules can help enhance signaling specificity and speed. Scaffold proteins are central components in neuronal synapses, where dynamic trafficking of synaptic proteins occurs. Mutations in scaffold proteins could have significant impact on synaptic structure and function. Postsynaptic density protein 95 (PSD95) is a member of the membrane-associated guanylate kinase (MAGUK) family of proteins and a scaffolding protein involved in the assembly and function of the postsynaptic density complex (1,2). SHANK proteins act as scaffolds at the neuronal post-synaptic density (PSD), where they play a critical role in PSD assembly of excitatory synapses during development (3,4). While recruitment of SHANK proteins to the synapse is independent of their interaction with Homer (5), proper synaptic targeting of SHANK1 is mediated by interactions between its PDZ domain and PSD proteins (6). Homer proteins (1-3) are scaffolds, composed of an EVH protein–binding domain, a coiled-coil domain and a leucine zipper domain. The EVH domain is a protein-protein binding module that binds to the proline-rich motifs of G-protein–coupled receptors (GPCRs), inositol 1,4,5-triphosphate (IP3) receptors (IP3Rs), ryanodine receptors, and TRP channels (7,8). The coiled-coil and the leucine zipper domains cause multimerization of Homers and assemble signaling proteins complexes. Spinophilin is a protein phosphatase 1 regulatory protein that interacts with a large number of proteins, including ion channel components and G-protein-coupled receptors (GPCRs). Spinophilin also interacts with actin filaments; phosphorylation of spinophilin at Ser94 and Ser177 disrupts this interaction (9,10).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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