Overview

  • Product name
  • Description
    Rabbit polyclonal to SNAP25
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chicken, Cow, Drosophila melanogaster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human SNAP25.

    Read Abcam's proprietary immunogen policy (Peptide available as ab41725.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Spinal Cord (Mouse) Tissue Lysate, Spinal Cord (Rat) Tissue lysate, Hippocampus (Mouse) Tissue Lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab41726 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
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).Can be blocked with Human SNAP25 peptide (ab41725).

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 (ab41726) at 1 µg/ml

    Lane 1 : Spinal Cord (Mouse) Tissue Lysate
    Lane 2 : Spinal Cord (Rat) Tissue lysate

    Lane 3 : Mouse brain tissue lysate - total protein (0 days) (ab7188)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

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

  • ICC/IF image of ab41726 stained human SH-SY5Y cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab41726, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • All lanes : Anti-SNAP25 antibody (ab41726) 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 Rabbit 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?

  • SNAP25 was immunoprecipitated using 0.5mg Rat Spinal Cord tissue lysate, 5µg of Rabbit polyclonal to SNAP25 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Rat Spinal Cord tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab41726.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 26kDa; SNAP25; non specific bands - 55 and 98kDa: We are unsure as to the identity of this extra band.

References

ab41726 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Non-reduced Denaturing (Extra PAGE ONE 5-20%)
Sample
Chicken Tissue lysate - whole (Brain and DRG)
Specification
Brain and DRG
Blocking step
Blocking one as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Feb 09 2015

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.

Read More
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.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No

Dr. Sophie Pezet

Verified customer

Submitted Mar 17 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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