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    snap25-antibody-ab41455.pdf

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Neuroscience Neurotransmission Secretory Vesicles SNAPs & SNAREs
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Anti-SNAP25 antibody (ab41455)

  • Datasheet
Reviews (2)Q&A (2)References (12)

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Immunocytochemistry - Anti-SNAP25 antibody (ab41455)
  • Western blot - Anti-SNAP25 antibody (ab41455)
  • Western blot - Anti-SNAP25 antibody (ab41455)
  • Immunocytochemistry - Anti-SNAP25 antibody (ab41455)
  • Western blot - Anti-SNAP25 antibody (ab41455)
  • Immunoprecipitation - Anti-SNAP25 antibody (ab41455)

Key features and details

  • Rabbit polyclonal to SNAP25
  • Suitable for: IP, ICC, WB
  • Reacts with: Mouse, Rat, Human, Zebrafish
  • Isotype: IgG

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Overview

  • Product name

    Anti-SNAP25 antibody
    See all SNAP25 primary antibodies
  • Description

    Rabbit polyclonal to SNAP25
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC
    Rat
    IP
    Rat
    WB
    Mouse
    Rat
    Human
    Zebrafish
    See all applications and species data
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human SNAP25.

    Read Abcam's proprietary immunogen policy (Peptide available as ab41724.)

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Secretory Vesicles
    • SNAPs & SNAREs
    • Neuroscience
    • Neurotransmission
    • Secretory Vesicles
    • Other
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • SNAPs & SNAREs
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Synapse marker
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Human SNAP25 peptide (ab41725)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Human SNAP25 protein (ab74529)
  • Recombinant Protein

    • Recombinant Human SNAP25 protein (ab74529)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab41455 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC
Rat
IP
Rat
WB
Mouse
Rat
Human
Zebrafish
All applications
Chicken
Cow
Application Abreviews Notes
IP
Use a concentration of 5 µg/ml.
ICC
Use a concentration of 1 - 5 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).
Notes
IP
Use a concentration of 5 µg/ml.
ICC
Use a concentration of 1 - 5 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 23 kDa).

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.
  • Target information above from: UniProt accession P60880 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 396444 Chicken
    • Entrez Gene: 540853 Cow
    • Entrez Gene: 6616 Human
    • Entrez Gene: 20614 Mouse
    • Entrez Gene: 25012 Rat
    • Entrez Gene: 30712 Zebrafish
    • Omim: 600322 Human
    • SwissProt: P60878 Chicken
    • SwissProt: Q17QQ3 Cow
    • SwissProt: P60880 Human
    • SwissProt: P60879 Mouse
    • SwissProt: P60881 Rat
    • SwissProt: Q5TZ66 Zebrafish
    • SwissProt: Q6PC54 Zebrafish
    • Unigene: 167317 Human
    • Unigene: 45953 Mouse
    • Unigene: 107689 Rat
    see all
  • 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
    • 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 antibody
    • Synaptosomal-associated protein, 25-KD antibody
    see all

Images

  • Immunocytochemistry - Anti-SNAP25 antibody (ab41455)
    Immunocytochemistry - Anti-SNAP25 antibody (ab41455)

    ab41455 staining SNAP25 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab41455 at 1µg/ml and ab192757, Mouse mono Anti-PSD95 antibody [K28/43] - Synaptic Marker. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

  • Western blot - Anti-SNAP25 antibody (ab41455)
    Western blot - Anti-SNAP25 antibody (ab41455)
    All lanes : Anti-SNAP25 antibody (ab41455) at 1 µg/ml

    Lane 1 : Spinal cord (Rat) tissue lysate
    Lane 2 : Hippocampus (Mouse) tissue Lysate

    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?

  • Western blot - Anti-SNAP25 antibody (ab41455)
    Western blot - Anti-SNAP25 antibody (ab41455)
    Anti-SNAP25 antibody (ab41455) at 1 µg/ml + Recombinant Human SNAP25 protein (ab74529) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa


    Exposure time: 1 minute
  • Immunocytochemistry - Anti-SNAP25 antibody (ab41455)
    Immunocytochemistry - Anti-SNAP25 antibody (ab41455)

    ab4455 staining SNAP25 in PC12 cells. The cells were fixed with 100% methanol (5 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 ab41455 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Western blot - Anti-SNAP25 antibody (ab41455)
    Western blot - Anti-SNAP25 antibody (ab41455)
    All lanes : Anti-SNAP25 antibody (ab41455) 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?


    Exposure time: 2 minutes
  • Immunoprecipitation - Anti-SNAP25 antibody (ab41455)
    Immunoprecipitation - Anti-SNAP25 antibody (ab41455)
    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 ab41455.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 26kDa; SNAP25

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (12)

    Publishing research using ab41455? Please let us know so that we can cite the reference in this datasheet.

    ab41455 has been referenced in 12 publications.

    • Comella-Bolla A  et al. Human Pluripotent Stem Cell-Derived Neurons Are Functionally Mature In Vitro and Integrate into the Mouse Striatum Following Transplantation. Mol Neurobiol 57:2766-2798 (2020). PubMed: 32356172
    • Wang X  et al. Long-term iron exposure causes widespread molecular alterations associated with memory impairment in mice. Food Chem Toxicol 130:242-252 (2019). PubMed: 31136779
    • Pan Y  et al. Pancreatic fibroblast growth factor 21 protects against type 2 diabetes in mice by promoting insulin expression and secretion in a PI3K/Akt signaling-dependent manner. J Cell Mol Med N/A:N/A (2018). PubMed: 30461198
    • Shirafuji T  et al. The Role of Cysteine String Protein a Phosphorylation at Serine 10 and 34 by Protein Kinase C? for Presynaptic Maintenance. J Neurosci 38:278-290 (2018). PubMed: 29167402
    • Reumann R  et al. The serine protease inhibitor neuroserpin is required for normal synaptic plasticity and regulates learning and social behavior. Learn Mem 24:650-659 (2017). PubMed: 29142062
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Flow Cytometry abreview for Anti-SNAP25 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Differentiated hNSCs)
    Permeabilization
    Yes - 0.25% Triton X-100 in DPBS
    Gating Strategy
    Undifferentiated Stem Cells (white)
    Specification
    Differentiated hNSCs
    Preparation
    Cell harvesting/tissue preparation method: Accutase
    Sample buffer: PBS
    Fixation
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 11 2015

    Question

    Thank you very much for your reply. To answer to your questions: 1) I use Broadman area 9, the same region that I used to quantify other synaptic protein (Munc 18, dynamin 1) and I had no problem with those. I find few papers reporting expression of SNAP-25 in BA9 from Human brain. 2) I used a classic blocking solution: PBS with 1% Tween-20 and 5% milk. 3) My lysis solution is 50mM Tris-HCl, 5mM EGTA, 10mM EDTA pH 7.4, so I have a crude homogenate of human brain samples (total protein) Let me know if this helps. I don’t mind having a replacement with the other antibody (ab41455), as soon as I can still ask for a refund if this one doesn’t work either....

    Read More

    Abcam community

    Verified customer

    Asked on Sep 14 2011

    Answer

    Thank you for your reply and for sending the information. I am sending a vial of ab41455 free of charge on the order ***, and this vial will also be covered by our Abpromise guarantee. If you have any problems with it, please let me know and I will be happy to arrange a refund for you. Let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Sep 14 2011

    Question

    Hello,   I contact you about an antibody ordered last January (req number ***): a Rabbit polyclonal antibody against SNAP25 (ref ab53723). I have been trying it few times now, on human and rat samples and I can’t get a good signal. I had a faint band for the rat cortex sample (see figure attached) and nothing for the human samples (n=3 samples from 3 different groups, 40µg and 60µg prot loaded), even with a high concentration of antibody (1/500) and a high intensity of exposure for the scan. When I stripped my membrane and run another wb for another Abcam antibody on the same samples (Dynamin-1; see bottom of the figure) I saw a good signal for all my rat and human samples. I am afraid the antibody SNAP25 (ref ab53723) doesn’t work on human brain samples, although it has been produced using a synthetic peptide derived from human SNAP25.   So I would like to have a refund for this antibody since I can’t use it on my samples. Also I would like to know if I could have a sample of your polyclonal antibody against SNAP25 (ref:ab41455) to try it on these samples?   I thank you in advance for your help.    

    Read More

    Abcam community

    Verified customer

    Asked on Sep 13 2011

    Answer

    Thank you for contacting us and for letting us know about the trouble with ab53723. I am sorry that the results have not been satisfactory. I do have a couple of questions so that I can better understand the situation: 1) What region of brain was the human sample taken from? SNAP25 is only expressed in certain regions which could account for the difference in signal from the rat cortex samples and the human brain samples. 2) What kind of blocking solution was used? 3) What lysis buffer was used to produce the tissue homogenate? We do guarantee all of our products to work as stated on the datasheet for up to 6 months after purchase, so I will be happy to either send a replacement such as ab41455 or refund the original purchase if the antibody is not working properly. I look forward to hearing from you. Please let me know if you have any questions.

    Read More

    Abcam Scientific Support

    Answered on Sep 13 2011

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-SNAP25 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Brain sections)
    Specification
    Brain sections
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    None
    Permeabilization
    No
    Read More

    Dr. Sophie Pezet

    Verified customer

    Submitted Mar 17 2009

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

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