Revision 1

#82760

Store at -20C

Silent Synapses Antibody Sampler Kit

1 Kit

(6 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes Product # Quantity Mol. Wt Isotype/Source
AMPA Receptor 1 (GluA1) (D4N9V) Rabbit Monoclonal Antibody1318520 µl100 kDaRabbit IgG
Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit Monoclonal Antibody7557420 µl100 kDaRabbit IgG
Phospho-AMPA Receptor 1 (GluA1) (Ser845) (D10G5) Rabbit Monoclonal Antibody808420 µl100 kDaRabbit IgG
AMPA Receptor 2 (GluA2) (E1L8U) Rabbit Monoclonal Antibody1360720 µl100 kDaRabbit IgG
PSD95 (D27E11) Rabbit Monoclonal Antibody345020 µl95 kDaRabbit IgG
NMDA Receptor 1 (GluN1) (D65B7) Rabbit Monoclonal Antibody570420 µl120 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from mouse brain, rat brain, and rat prefrontal cortex tissues using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb.
undefined Image 1: Silent Synapses Antibody Sampler Kit
Western blot analysis of extracts from mouse brain, rat brain, and human cortex tissues using AMPA Receptor 2 (GluA2) (E1L8U) Rabbit mAb.
undefined Image 2: Silent Synapses Antibody Sampler Kit
Western blot analysis of extracts from human cerebellum and rat brain using PSD95 (D27E11) XP® Rabbit mAb.
undefined Image 3: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from rat and mouse brain using NMDA Receptor1 (GluN1) (D65B7) Rabbit mAb.
undefined Image 4: Silent Synapses Antibody Sampler Kit
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.
undefined Image 5: Silent Synapses Antibody Sampler Kit
Simple WesternTM analysis of lysates (0.1 mg/mL) from Mouse Brain cells using Phospho-AMPA Receptor 1 (GluA1) (Ser831) (A5O2P) Rabbit mAb #75574. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 6: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 7: Silent Synapses Antibody Sampler Kit

Simple WesternTM analysis of lysates (0.1 mg/mL) from Mouse Brain cells using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb #13185. 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 JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

undefined Image 8: Silent Synapses Antibody Sampler Kit
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).
undefined Image 9: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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).
undefined Image 10: Silent Synapses Antibody Sampler Kit
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.
undefined Image 11: Silent Synapses Antibody Sampler Kit
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.
undefined Image 12: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 13: Silent Synapses Antibody Sampler Kit
Confocal immunofluorescent analysis of mouse hippocampus using AMPA Receptor 1 (GluA1) (D4N9V) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 14: Silent Synapses Antibody Sampler Kit
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.
undefined Image 15: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of human iPSC-derived cortical glutamatergic neurons at 7 days in vitro using PSD95 (D27E11) XP® Rabbit mAb (green), β3-Tubulin (E9F3E) Mouse mAb #45058 (red), and DAPI #4083 (blue). iCell GlutaNeurons were kindly provided by FUJIFILM Cellular Dynamics, Inc.
undefined Image 16: Silent Synapses Antibody Sampler Kit
Immunoprecipitation of Phospho-AMPA Receptor 1 (GluA1) (Ser845) from mouse brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-AMPA Receptor 1 (GluA1) (Ser845) (D10G5) Rabbit mAb. Western blot analysis was performed using Phospho-AMPA Receptor 1 (GluA1) (Ser845) (D10G5) Rabbit mAb.
undefined Image 17: Silent Synapses Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.