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82760
Silent Synapses Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Silent Synapses Antibody Sampler Kit #82760

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Silent Synapses Antibody Sampler Kit: Image 1

Confocal immunofluorescent analysis of mouse hippocampus using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Silent Synapses Antibody Sampler Kit: Image 2

Western blot analysis of extracts from mouse brain, untreated (-) or λ-phosphatase-treated (+), using Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit mAb (upper) and AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb #13185 (lower).

Silent Synapses Antibody Sampler Kit: Image 3

Western blot analysis of extracts from mouse brain and rat brain using Phospho-AMPA Receptor 1 (GluA1) (Ser845) (D10G5) Rabbit mAb. The phospho-specificity of the antibody was verified by blocking with a phospho or nonphosphopeptide.

Silent Synapses Antibody Sampler Kit: Image 4

Immunoprecipitation of AMPA Receptor 2 (GluA2) from rat brain extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or AMPA Receptor 2 (GluA2) (E1L8U) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using AMPA Receptor 2 (GluA2) (E1L8U) Rabbit mAb.

Silent Synapses Antibody Sampler Kit: Image 5

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).

Silent Synapses Antibody Sampler Kit: Image 6

Immunoprecipitation of NMDA Receptor 1 (GluN1) protein from adult rat brain extracts. Lane 1 is NMDA Receptor 1 (GluN1) (D65B7) Rabbit mAb, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is 10% input. Western blot analysis was performed using NMDA Receptor 1 (GluN1) (D65B7) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.

Silent Synapses Antibody Sampler Kit: Image 7

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.

Silent Synapses Antibody Sampler Kit: Image 8

Immunoprecipitation of AMPA Receptor 1 (GluA1) from mouse brain extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb.

Silent Synapses Antibody Sampler Kit: Image 9

Western blot analysis of extracts from mouse brain, rat brain, and human cortex tissues using AMPA Receptor 2 (GluA2) (E1L8U) Rabbit mAb.

Silent Synapses Antibody Sampler Kit: Image 10

Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.

Silent Synapses Antibody Sampler Kit: Image 11

Western blot analysis of extracts from rat and mouse brain using NMDA Receptor1 (GluN1) (D65B7) Rabbit mAb.

Silent Synapses Antibody Sampler Kit: Image 12

Western blot analysis of extracts from mouse brain, rat brain, and rat prefrontal cortex tissues using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb.

To Purchase # 82760T
Product # Size Price
82760T
1 Kit  (6 x 20 µl) $ 445

Product Description

The Silent Synapses Antibody Sampler Kit provides an economical means of detecting the activation of AMPA-type glutamate receptors (AMPAR) using phospho-specific and control antibodies. AMPARs expression can be compared to other synaptic components including NMDA-type glutamate receptor subunit GluN1 and the synaptic scaffolding protein PSD95. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Silent Synapses Antibody Sampler Kit detects its target protein at endogenous levels. The phospho-specific antibodies recognize

human AMPA Receptor 1 (GluA1) only when phosphorylated at the indicated residues. While the literature refers to the GluA1 phospho-residues as Ser831 and Ser845, the corresponding residues for UniProt ID #P42261 are Ser849 and Ser863, respectively.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala275 of human AMPA Receptor 1 (GluA1) protein, Ser52 of human AMPA Receptor 2 (GluA2) protein, Gln53 of human PSD95, and Pro660 of the human NMDA Receptor 1 (GluN1) protein. Activation-state specific monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser831 and Ser845 of human AMPA Receptor 1 (GluA1) protein.

Background

AMPA- (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid), kainate-, and NMDA- (N-methyl-D-aspartate) receptors are the three main families of ionotropic glutamate-gated ion channels. AMPA receptors (AMPARs) are composed of four subunits (GluA1-4), which assemble as homo- or hetero-tetramers to mediate the majority of fast excitatory transmissions in the central nervous system. AMPARs are implicated in synapse formation, stabilization, and plasticity (1). In contrast to GluA2-containing AMPARs, AMPARs that lack GluA2 are permeable to calcium (2). Post-transcriptional modifications (alternative splicing, nuclear RNA editing) and post-translational modifications (glycosylation, phosphorylation) result in a very large number of permutations, fine-tuning the kinetic properties and surface expression of AMPARs representing key pathways to mediate synaptic plasticity (3). During development and mature states, some synapses exhibit “silent synapses” that lack functional AMPAR-mediated transmission. Synapses become “unsilenced” by post-translational modification of GluAs, particularly GluA1, which alters its kinetic properties and/or surface expression while other synaptic components, such as other glutamate receptors like NMDARs and postsynaptic scaffolding proteins like PSD95, remain unaltered. Conversely, reducing the AMPAR kinetic properties and surface expression can silence synapses. Key post-translational modifications implicated in regulating these processes include phosphorylation of GluA1 at Ser831 and Ser845 (4). Research studies have implicated activity-dependent changes in AMPARs in a variety of diseases, including Alzheimer’s, amyotrophic lateral sclerosis (ALS), stroke, and epilepsy (1).

  1. Palmer, C.L. et al. (2005) Pharmacol Rev 57, 253-77.
  2. Cull-Candy, S. et al. (2006) Curr Opin Neurobiol 16, 288-97.
  3. Huganir, R.L. and Nicoll, R.A. (2013) Neuron 80, 704-17.
  4. Diering, G.H. et al. (2016) Proc Natl Acad Sci U S A 113, E4920-7.

Pathways & Proteins

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