Overview

  • Product name

    Anti-SHANK3 antibody [N69/46]
    See all SHANK3 primary antibodies
  • Description

    Mouse monoclonal [N69/46] to SHANK3
  • Host species

    Mouse
  • Specificity

    No cross-reactivity with SHANK1 or SHANK2.
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Cow
  • Immunogen

    Synthetic peptide:

    PEKLPGSLRKGIPRTKSV

    , corresponding to internal sequence amino acids 840 - 857 of Rat SHANK3 (Acc. No. Q9JLU4).

  • General notes

    The clone number has been updated from S69-46 to N69/46, both clone numbers name the same antibody clone.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab93607 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 190 kDa (predicted molecular weight: 186 kDa).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    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.
  • Tissue specificity

    Expressed in the cerebral cortex and the cerebellum.
  • Involvement in disease

    Note=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.
    Note=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) [MIM:613950]. 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.
  • Sequence similarities

    Contains 6 ANK repeats.
    Contains 1 PDZ (DHR) domain.
    Contains 1 SAM (sterile alpha motif) domain.
    Contains 1 SH3 domain.
  • Cellular localization

    Cytoplasm. Cell junction > synapse. Cell junction > synapse > postsynaptic cell membrane > postsynaptic density. Postsynaptic density of neuronal cells.
  • Information by UniProt
  • Database links

  • Alternative names

    • AI841104 antibody
    • DEL22q13.3 antibody
    • KIAA1650 antibody
    • Proline rich synapse associated protein 2 antibody
    • Proline-rich synapse-associated protein 2 antibody
    • ProSAP2 antibody
    • PSAP2 antibody
    • SH3 and multiple ankyrin repeat domains 3 antibody
    • SH3 and multiple ankyrin repeat domains protein 3 antibody
    • SH3/ankyrin domain gene 3 antibody
    • SHAN3_HUMAN antibody
    • Shank postsynaptic density protein antibody
    • Shank3 antibody
    • Shank3b antibody
    • SPANK 2 antibody
    • SPANK2 antibody
    see all

Images

  • Anti-SHANK3 antibody [N69/46] (ab93607) at 1/1000 dilution + Rat brain membrane lysate at 15 µg

    Secondary
    Sheep Anti-Mouse IgG-HRP

    Predicted band size: 186 kDa

  • Immunohistochemical analysis of mouse backskin tissue sections labling SHANK3 with ab93607 at 1/100 dilution. FITC Goat anti-mouse at 1/50 was used as the secondary antibody. 

     

  • Overlay histogram showing SH-SY5Y cells stained with ab93607 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab93607, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Zhang Y  et al. Effect of intraperitoneal or intracerebroventricular injection of streptozotocin on learning and memory in mice. Exp Ther Med 16:2375-2380 (2018). Read more (PubMed: 30186480) »
See 1 Publication for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer

Thank you for confirming these details.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your reply.

I am happy to send this out as soon as possible. Can you confirm your previous order number for ab93607? I can't seem to locate it in our system and need either the order number or PO to arrange for the replacement to be sent.


I look forward to your reply so that I may place this order for you.

Read More

Answer

Thank you for your reply.


I would like to suggest ab104702. There is a short lead time, but it is validated in Rat and WB. Please let me know if you would like to receive this antibody as a replacement.


I look forward to your reply so that I may process your request.

Read More

Answer

Thank you for your reply.


In searching for a protocol for ab80983, I have confirmed that this product was tested with the recombinant protein used as the immunogen, but not with endogenous protein. Therefore, I do not have an optimized protocol. I have several other antibodies available - can you confirm the species of your sample? I would be happy to recommend another product, which would be guaranteed by the Abpromise should it not work as described on the datasheet.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact me if you have any other questions.

Read More

Answer

Thank you for your reply.


I am sorry that you are having difficulties with this antibody in WB. Looking at your blot, it seems that the 1:500 (5ug/ml)dilution is producing a faint band at the predicted molecular weight and using a higher dilution may help with this (we recommend up to 10ug/ml, 1:100).


Alternatively, I would be happy to suggest an alternative SHANK3 antibody. For example, ab80983 is validated for WB in human, mouse, and rat samples. Unfortunately, none of the other antibodies to this target are specifically validated for IP. The information we have on ab93607 was collected from customer feedback and we do not have any additional information.


Please let me know if you are interested in receiving ab80983 as a replacement. If you are concerned about using the antibody in IP, then I am happy to offer a credit instead. Please let me know which you would prefer.


I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

Read More

Answer

Thank you for your reply.

I am sorry that you are experiencing difficulties with this antibody in WB. Below, please find the protocol used with this antibody.


1. Transfer and immobilize antigen on nitrocellulose or PVDF membrane.


2. Block by immersing the membrane in TTBS + 1% BSA. Use just enough solution to cover the membrane.


3. Transfer the membrane to diluted solution of primary antibody in TTBS + 1% BSA. The appropriate dilution should be determined by trial and error but we recommend testing between 1-10ug/ml of antibody. Incubate overnight at 4oC.


4. Wash with 3 changes of TTBS for 5 minutes each with gentle agitation.


5. Transfer the membrane to a dilute solution of biotinylated secondary antibody in TTBS. . Incubate for 30 minutes with gentle agitation.


6. Repeat step 4.


7. Transfer the membrane to a solution of peroxidase conjugated streptavidin appropriately diluted in TTBS. Incubate at 30 minutes with gentle agitation.


8. During the above incubation, prepare the enzyme substrate. Use DAB for a dark brown color or TMB for a bright blue color.


9. Wash as in step 4.


10. Transfer the membrane to the substrate solution. Incubate until color develops sufficiently (usually 2-20 minutes).


11. Wash with 2 changes of water for 5 minutes each with gentle agitation. Allow membrane to dry and store in the dark.


I hope this information is helpful. If these modifications do not improve your results with this antibody I am happy to offer a replacement or credit.

Read More

Question
Answer

Thank you for contacting us.

The product was purified from acites (ascitic fluid).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for contacting Abcam. I have attached a WB image demonstrating the specificity of this antibody. It's a western in brain membrane lysates using a 1:1000 dilution. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up