Anti-SNAP25 antibody (ab66066)

Mouse monoclonal SNAP25 antibody. Validated in WB, ELISA, Flow Cyt, ICC/IF and tested in Mouse, Human, Zebrafish. Cited in 3 publication(s). Immunogen corresponding to recombinant fragment.

Overview

  • Product name
  • Description
    Mouse monoclonal to SNAP25
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, WB, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Rat, Chicken, Dog, Xenopus laevis, Cynomolgus monkey, Opossum, Orangutan
  • Immunogen

    Recombinant tagged fragment: RMLQLVEESK DAGIRTLVML DEQGEQLDRV EEGMNHINQD MKEAEKNLKD LGKCCGLFIC PCNKLKSSDA YKKAWGNNQD GVVASQPARV VDEREQMAIS, corresponding to amino acids 31-130 of Human SNAP25

  • Positive control
    • SNAP25 transfected 293T cell lysate. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: SKNSH.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 4th April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab66066 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
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    t-SNARE involved in the molecular regulation of neurotransmitter release. May play an important role in the synaptic function of specific neuronal systems. Associates with proteins involved in vesicle docking and membrane fusion. Regulates plasma membrane recycling through its interaction with CENPF.
  • Tissue specificity
    Neurons of the neocortex, hippocampus, piriform cortex, anterior thalamic nuclei, pontine nuclei, and granule cells of the cerebellum.
  • Sequence similarities
    Belongs to the SNAP-25 family.
    Contains 2 t-SNARE coiled-coil homology domains.
  • Post-translational
    modifications
    Palmitoylated. Cys-85 appears to be the main site, and palmitoylation is required for membrane association.
  • Cellular localization
    Cytoplasm > perinuclear region. Cell membrane. Cell junction > synapse > synaptosome. Membrane association requires palmitoylation. Expressed throughout cytoplasm, concentrating at the perinuclear region.
  • Information by UniProt
  • Database links
  • Alternative names
    • bA416N4.2 antibody
    • Bdr antibody
    • CMS18 antibody
    • dJ1068F16.2 antibody
    • FLJ23079 antibody
    • HGNC:11132 antibody
    • MGC105414 antibody
    • MGC139754 antibody
    • Resistance to inhibitors of cholinesterase 4 homolog antibody
    • RIC 4 antibody
    • RIC4 antibody
    • SEC 9 antibody
    • SEC9 antibody
    • SNAP 25 antibody
    • SNAP antibody
    • SNAP-25 antibody
    • SNAP-25B antibody
    • SNAP25 antibody
    • SNP 25 antibody
    • SNP25 antibody
    • SNP25_HUMAN antibody
    • sp antibody
    • SUP antibody
    • Super protein antibody
    • Synaptosomal associated 25 kDa protein antibody
    • Synaptosomal associated protein antibody
    • Synaptosomal associated protein 25 antibody
    • Synaptosomal associated protein 25kDa antibody
    • Synaptosomal-associated 25 kDa protein antibody
    • Synaptosomal-associated protein 25 antibody
    • Synaptosomal-associated protein, 25-KD antibody
    see all

Images

  • All lanes : Anti-SNAP25 antibody (ab66066) at 1 µg/ml

    Lane 1 : SNAP25 transfected 293T cell lysate
    Lane 2 : Non-transfected 293T cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG (H&L)-HRP at 1/2500 dilution

    Predicted band size: 23 kDa
    Observed band size: 27 kDa
    why is the actual band size different from the predicted?



    This image was generated using the ascites version of the product.

  • All lanes : Anti-SNAP25 antibody (ab66066) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : SH-SY5Y (Human neuroblastoma cell line) whole cell lysate at 20 µg
    Lane 4 : Mouse brain homogenate at 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    This image was generated using the ascites version of the product.

  • Overlay histogram showing SH-SY5Y cells stained with ab66066 (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 followed by the antibody (ab66066, 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 IgG1 [ICIGG1] (ab91353, 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.

    This image was generated using the ascites version of the product.

  • ab66066 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab66066 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:
  • Mo J  et al. PRRT2 deficiency induces paroxysmal kinesigenic dyskinesia by influencing synaptic function in the primary motor cortex of rats. Neurobiol Dis 121:274-285 (2019). Read more (PubMed: 30347267) »
  • Yang H  et al. Brain-Specific SNAP-25 Deletion Leads to Elevated Extracellular Glutamate Level and Schizophrenia-Like Behavior in Mice. Neural Plast 2017:4526417 (2017). Read more (PubMed: 29318050) »
See all 3 Publications for this product

Customer reviews and Q&As

Question

I'm working on a project that involves developing a procedure to map epitopes using mass spec. To do this we are using different types of anitbodies specific to SNAP25. The SNAP25 we are using is Genway,human recombinant SNAP25 made in E. coli. Here is a link to the protein and all of the specifications: http://www.genwaybio.com/product_info.php?products_id=46229 I have already mapped a couple of epitopes including abcam's 66066. What I am looking for is a anti-SNAP25 antibody that has an epitope in a region of SNAP25 that we have had success in dectecting with the mass spec. These areas include: -(43)MQRRADQ LADESLESTR RMLQLVEESK DAGIRTLVML DEQGEQLERI EEGMDQINK(98)- -(150)VDEREQMAIS GGFI(163)- -(170)ARENEMDENL EQVSGIIGNL RHMALDMGNE IDTQNRQIDR IME(203)- Also I have already used an antibody with the epitope of -(214)ANQRATKMLGSG(226)- so I would not be interested in another antibody in the same region. In conclusion I am looking for an anti-SNAP25 anitbody who's epitope is somewhere in the region(s) above. If you do find an antibody it would be helpful if you could include the sequence of the epitope. It is important to the experiment to know the exact sequence. I really hope this email is not too confusing. Also this is my second time writing, the first time I got an email back with specific questions but I could not reply back to the person trying to help me. Not sure what the problem was, but I think I have included all the info you may need. Thanks for your help, have a good day.

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Answer

Thank you for providing the additional information. I see your earlier e-mail in our records. We have 15 SNAP25 antibodies in our catalogue. Some are polyclonals raised against full-length SNAP25, and since these will potentially recognize multiple epitopes along the entire protein, they will not be useful for your project. Similarly, some others are monoclonal antibodies raised against the full-length protein but the single epitope they recognize has not been mapped to a specific range of amino acids. Of the remaining antibodies, several are raised agains the C-terminus which you already have an antibody against. One exception: Antibody ab41726 is raised agianst a peptide (amino acids ASQPARVVDEREQMA)which has some overlap with a region you specified: (150)VDEREQMAISGGFI(163) However, the antibody may recognize an epitope composed of amino acids outside the range: ASQPARV. There are several other antibodies which may be sueful but I need to obtain the immunogen sequences from the laboratories that produce them: ab53723 ab109105 ab68859 ab72396 ab68858 ab72394 I will forward the information when I receive it. Hopefully one or more of these will meet your needs.

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