Overview

  • Product name
  • Description
    Rabbit polyclonal to SNAP25
  • Tested applications
    Suitable for: IP, ICC/IF, WB, ICC, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Guinea pig, Cow, Zebrafish
    Predicted to work with: Human, Chimpanzee, Rhesus monkey, Goldfish
  • Immunogen

    Synthetic peptide corresponding to Mouse SNAP25 aa 195-206.
    Sequence:

    ANQRATKMLGSG


    (Peptide available as ab5843)

  • Positive control
    • ICC/IF: PC12 cell line. WB: Whole rat brain extract.

Properties

Applications

Our Abpromise guarantee covers the use of ab5666 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 1/100.

Immunofluorescence staining of SNAP-25 in cultured rat hippocampal cells with ab5666 yields a pattern consistent with cytoplasmic and plasma membrane staining.

WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 25 kDa.Can be blocked with SNAP25 peptide (ab5843).
ICC Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 3-5µg for 106 cells.

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

Anti-SNAP25 antibody images

  • All lanes : Anti-SNAP25 antibody (ab5666)

    Lane 1 : Mouse cerebellum
    Lane 2 : Mouse brain
    Lane 3 : Rat brain
    Lane 4 : Membrane enriched extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate
  • ICC/IF image of ab5666 stained PC12 cells. The cells were 4% paraformaldehyde 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 (ab5666, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • Flow cytometry analysis of Neuro-2a cells using ab5666.Cells were fixed with 70% ethanol for 10 minutes,permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature.Cells were labeled with SNAP25 (red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA.After incubation at room temperature for 2 hours,the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody at a dilution of 1:400 for 30 minutes at room temperature.The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

  • SNAP25 was immunoprecipitated using 0.5mg Mouse Brain 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, Mouse Brain 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 ab5666.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 25kDa; SNAP25
  • Immunlocalization of SNAP-25 in cultured rat hippocampal cells using ab5666.
  • Anti-SNAP25 antibody (ab5666) at 1/10000 dilution + Whole tissue lysate prepared from mouse hippocampus.

    Secondary
    HRP conjugated goat anti-rabbit Ig. at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 25 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-SNAP25 antibody (ab5666)

This product has been referenced in:
  • Niere F  et al. Analysis of Proteins That Rapidly Change Upon Mechanistic/Mammalian Target of Rapamycin Complex 1 (mTORC1) Repression Identifies Parkinson Protein 7 (PARK7) as a Novel Protein Aberrantly Expressed in Tuberous Sclerosis Complex (TSC). Mol Cell Proteomics 15:426-44 (2016). Read more (PubMed: 26419955) »
  • Radford H  et al. PERK inhibition prevents tau-mediated neurodegeneration in a mouse model of frontotemporal dementia. Acta Neuropathol 130:633-42 (2015). WB . Read more (PubMed: 26450683) »

See all 13 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted Feb 09 2015

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Sample
Rat Tissue lysate - whole (hippocampus,tissue lysate)
Specification
hippocampus,tissue lysate
Treatment
RIPA
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C
Username

Abcam user community

Verified customer

Submitted Jun 06 2014

Thank you for your recent telephone enquiry.

I can confirm that ab5666 SNAP25 antibodyis provided at 1 mg/ml. I have updated the online datasheet.

I hope this will be helpful to you. If you have any further questions, please do no...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (hippocampus)
Specification
hippocampus
Gel Running Conditions
Reduced Denaturing (12% TGS)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 20 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain)
Loading amount
100 µg
Specification
brain
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Nov 17 2006

I'm very sorry to hear you are experiencing problems with ab5666 in both WB and IHC, this antibody has not been tested on sections (only in cell culture) and without a detailed IHC protocol it is hard to determine what the problem may be in that applic...

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