Western Blot (WB)
(Figure 1. Western blot analysis of SHANK3 using anti-SHANK3 antibody (MBS1753413).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-SHANK3 antigen affinity purified polyclonal antibody (Catalog # MBS1753413) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for SHANK3 at approximately 180-190KD. The expected band size for SHANK3 is at 185KD. )
Rabbit SHANK3 Polyclonal Antibody | anti-SHANK3 antibody
Anti-SHANK3 Antibody
IHC-P: 2-5ug/ml|Human, Rat|
FC/FACS/FCM: 1-3ug/1x106 cells|Human, Mouse, Rat|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of SHANK3 using anti-SHANK3 antibody (MBS1753413).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-SHANK3 antigen affinity purified polyclonal antibody (Catalog # MBS1753413) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for SHANK3 at approximately 180-190KD. The expected band size for SHANK3 is at 185KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of SHANK3 using anti-SHANK3 antibody (MBS1753413).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat brain tissue lysatesLane 2: mouse brain tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-SHANK3 antigen affinity purified polyclonal antibody (Catalog # MBS1753413) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for SHANK3 at approximately 180-190KD. The expected band size for SHANK3 is at 185KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of SHANK3 using anti SHANK3 antibody (MBS1753413).SHANK3 was detected in paraffin-embedded section of human meningoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-SHANK3 Antibody (MBS1753413) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of SHANK3 using anti SHANK3 antibody (MBS1753413).SHANK3 was detected in paraffin-embedded section of human meningoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-SHANK3 Antibody (MBS1753413) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of SHANK3 using anti SHANK3 antibody (MBS1753413).SHANK3 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-SHANK3 Antibody (MBS1753413) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of SHANK3 using anti SHANK3 antibody (MBS1753413).SHANK3 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-SHANK3 Antibody (MBS1753413) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of Raji cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing Raji cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of Raji cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing Raji cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of HEPA1-6 cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing HEPA1-6 cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 5. Flow Cytometry analysis of HEPA1-6 cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing HEPA1-6 cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of NRK cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing NRK cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 6. Flow Cytometry analysis of NRK cells using anti-SHANK3 antibody (MBS1753413).Overlay histogram showing NRK cells stained with MBS1753413 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SHANK3 Antibody (MBS1753413,1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Boeckers TM, Bockmann J, Kreutz MR, Gundelfinger ED (2002). "ProSAP/Shank proteins - a family of higher order organizing molecules of the postsynaptic density with an emerging role in human neurological disease". J. Neurochem. 81 (5): 903-10.
NCBI and Uniprot Product Information
NCBI Description
This gene is a member of the Shank gene family. Shank proteins are multidomain scaffold proteins of the postsynaptic density that connect neurotransmitter receptors, ion channels, and other membrane proteins to the actin cytoskeleton and G-protein-coupled signaling pathways. Shank proteins also play a role in synapse formation and dendritic spine maturation. Mutations in this gene are a cause of autism spectrum disorder (ASD), which is characterized by impairments in social interaction and communication, and restricted behavioral patterns and interests. Mutations in this gene also cause schizophrenia type 15, and are a major causative factor in the neurological symptoms of 22q13.3 deletion syndrome, which is also known as Phelan-McDermid syndrome. Additional isoforms have been described for this gene but they have not yet been experimentally verified. [provided by RefSeq, Mar 2012]
Uniprot Description
SHANK3: Seems to be an adapter protein in the postsynaptic density (PSD) of excitatory synapses that 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. May play a role in the structural and functional organization of the dendritic spine and synaptic junction. A chromosomal aberration involving SHANK3 is found in patients with chromosome 22q13.3 deletion syndrome. Translocation t(12;22)(q24.1;q13.3) with APPL2/DIP13B. Defects in SHANK3 are associated with autism spectrum disorders (ASD). ASD are characterized by impairments in reciprocal social interaction and communication as well as restricted and stereotyped patterns of interest and activities. ASD include forms with moderate to severe cognitive impairment and milder forms with higher cognitive ability (Asperger syndrome). Defects in SHANK3 are the cause of schizophrenia type 15 (SCZD15). SCZD15 is a complex, multifactorial psychotic disorder or group of disorders characterized by disturbances in the form and content of thought (e.g. delusions, hallucinations), in mood (e.g. inappropriate affect), in sense of self and relationship to the external world (e.g. loss of ego boundaries, withdrawal), and in behavior (e.g bizarre or apparently purposeless behavior). Although it affects emotions, it is distinguished from mood disorders in which such disturbances are primary. Similarly, there may be mild impairment of cognitive function, and it is distinguished from the dementias in which disturbed cognitive function is considered primary. Some patients manifest schizophrenic as well as bipolar disorder symptoms and are often given the diagnosis of schizoaffective disorder. 2 isoforms of the human protein are produced by alternative splicing.
Protein type: Adaptor/scaffold
Chromosomal Location of Human Ortholog: 22q13.3
Cellular Component: postsynaptic membrane; neuron projection; cytoplasm; plasma membrane; dendritic spine; cell junction
Molecular Function: protein C-terminus binding; identical protein binding; ionotropic glutamate receptor binding; protein binding; zinc ion binding; GKAP/Homer scaffold activity; actin binding; SH3 domain binding
Biological Process: negative regulation of actin filament bundle formation; MAPKKK cascade; vocal learning; social behavior; learning; positive regulation of synaptic transmission, glutamatergic; positive regulation of long-term neuronal synaptic plasticity; memory; positive regulation of synapse structural plasticity; synaptogenesis; striatal medium spiny neuron differentiation; adult behavior; brain morphogenesis; negative regulation of cell volume
Disease: Schizophrenia 15; Phelan-mcdermid Syndrome
Research Articles on SHANK3
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Product Notes
The SHANK3 shank3 (Catalog #AAA1753413) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-SHANK3 Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's SHANK3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.1-0.25ug/ml|Mouse, Rat| IHC-P: 2-5ug/ml|Human, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Human, Mouse, Rat| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the SHANK3 shank3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "SHANK3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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