Anti-SHANK1/SHANK3 Antibody (56578)

Host Quantity Applications Species Reactivity Data Sheet  
Mouse 100ug WB,IHC,ICC/IF Human, Mouse, Rat
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Overview
Product Name Anti-SHANK1/SHANK3 Antibody (56578)
Description Anti-SHANK1/SHANK3 Mouse Monoclonal Antibody
Target SHANK1/SHANK3
Species Reactivity Human, Mouse, Rat
Applications WB,IHC,ICC/IF
Host Mouse
Clonality Monoclonal
Clone ID S367-51
Isotype IgG2a
Immunogen Fusion protein corresponding to aa 538-626 (SH3 domain) of rat SHANK3. This sequence is 100
Properties
Form Liquid
Concentration 1.0 mg/mL
Formulation PBS, pH 7.4, 0.1% sodium azide, 50% glycerol.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Buffer Anti-Microbial 0.1% Sodium Azide
Buffer Cryopreservative 50% Glycerol
Format Purified
Purification Purified by Protein G affinity chromatography
Specificity Information
Specificity This antibody recognizes human, mouse, and rat SHANK1/SHANK3.
Target Name SH3 and multiple ankyrin repeat domains protein 3
Target ID SHANK1/SHANK3
Uniprot ID Q9JLU4
Alternative Names Shank3, Proline-rich synapse-associated protein 2, ProSAP2, SPANK-2
Gene Name Shank3
Sequence Location Cytoplasm. Cell junction, synapse, postsynaptic density. Cell projection, dendritic spine. Note=In neuronal cells, extends into the region subjacent to the PSD.
Biological Function Major scaffold postsynaptic density protein which interacts with multiple proteins and complexes to orchestrate the dendritic spine and synapse formation, maturation and maintenance. Interconnects receptors of the postsynaptic membrane including NMDA-type and metabotropic glutamate receptors via complexes with GKAP/PSD-95 and HOMER, respectively, and the actin-based cytoskeleton. Plays a role in the structural and functional organization of the dendritic spine and synaptic junction through the interaction with Arp2/3 and WAVE1 complex as well as the promotion of the F-actin clusters. By way of this control of actin dynamics, participates in the regulation of developing neurons growth cone motility and the NMDA receptor-signaling. Also modulates GRIA1 exocytosis and GRM5/MGLUR5 expression and signaling to control the AMPA and metabotropic glutamate receptor-mediated synaptic transmission and plasticity. May be required at an early stage of synapse formation and be inhibited by IGF1 to promote synapse maturation. {PubMed:21606927, PubMed:21795692, PubMed:23100419, PubMed:23897824, PubMed:24089484}.
Research Areas Neuroscience
Background SHANK proteins are scaffolding adaptors that have been shown to integrate neurotransmitter receptors into the cortical cytoskeleton at postsynaptic densities. SHANK1-3 of the SHANK/ProSAP family are molecular scaffolds in the postsynaptic density (PSD). SHANK recruits betaPIX and PAK to dendritic spines to regulate postsynaptic structure and interacts with ionotropic receptor and metabotropic glutamate receptor complexes. Transcript splice variation in the Shank family influences the spectrum of Shank-interacting proteins in the PSDs of adult and developing brain to ensure normal development.
Application Images
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody, Clone S367-51 (56578). Tissue: Neuroblastoma cells (SH-SY5Y). Species: Human. Fixation: 4% PFA for 15 min. Primary Antibody: Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody (56578) at 1:100 for overnight at 4°C with slow rocking. Secondary Antibody: AlexaFluor 488 at 1:1000 for 1 hour at RT. Counterstain: Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 min at RT. (A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) SHANK1/SHANK3 Antibody (D) Composite.
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody, Clone S367-51 (56578). Tissue: Neuroblastoma cell line (SK-N-BE). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody (56578) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000; 1:5000 for 60 min RT, 5 min RT. Localization: Cytoplasm, Nucleus. Magnification: 60X. (A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) SHANK1/SHANK3 Antibody. (D) Composite.
Description Western Blot analysis of Monkey COS cells transfected with HA-tagged Shank1 showing detection of ~190 kDa SHANK1/SHANK3 protein using Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody, Clone S367-51 (56578). Lane 1: Molecular Weight Ladder. Lane 2: Monkey COS cells transfected with HA-tagged Shank1. Load: 15 µg. Block: 2% BSA and 2% Skim Milk in 1X TBST. Primary Antibody: Mouse Anti-SHANK1/SHANK3 Monoclonal Antibody (56578) at 1:200 for 16 hours at 4°C. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:1000 for 1 hour RT. Color Development: ECL solution for 6 min in RT. Predicted/Observed Size: ~190 kDa.
Additional Information
Handling
Storage This product is stable for at least one (1) year at -20°C.
Dilution Instructions Dilute in PBS or medium that is identical to that used in the assay system.
Application Instructions

Immunofluorescence: use at 10ug/ml.

These are recommended concentrations.

Endusers should determine optimal concentrations for their application.
References & Data Sheet