Anti-SNAP25 抗体 (ab5666)

Reviews (4)Q&A (2)References (33)

Key features and details

  • Rabbit polyclonal to SNAP25
  • Suitable for: ICC, WB, Flow Cyt
  • Reacts with: Mouse, Rat, Chicken, Guinea pig, Cow, Zebrafish
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-SNAP25 antibody
    SNAP25 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to SNAP25

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: ICC, WB, Flow Cytmore details

  • 種交差性

    交差種: Mouse, Rat, Chicken, Guinea pig, Cow, Zebrafish
    交差が予測される動物種: Human, Chimpanzee, Rhesus monkey, Goldfish

  • 免疫原

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

    ANQRATKMLGSG


    (Peptide available as ab5843)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
ICC Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 25 kDa.
Flow Cyt Use 3-5µg for 106 cells.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    Neurons of the neocortex, hippocampus, piriform cortex, anterior thalamic nuclei, pontine nuclei, and granule cells of the cerebellum.

  • 配列類似性

    Belongs to the SNAP-25 family.
    Contains 2 t-SNARE coiled-coil homology domains.

  • 翻訳後修飾

    Palmitoylated. Cys-85 appears to be the main site, and palmitoylation is required for membrane association.

  • 細胞内局在

    Cytoplasm > perinuclear region. Cell membrane. Cell junction > synapse > synaptosome. Membrane association requires palmitoylation. Expressed throughout cytoplasm, concentrating at the perinuclear region.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-SNAP25 antibody (ab5666)
    Western blot - Anti-SNAP25 antibody (ab5666)
    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
    All lanes : Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate
  • Immunocytochemistry - Anti-SNAP25 antibody (ab5666)
    Immunocytochemistry - Anti-SNAP25 antibody (ab5666)
    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 - Anti-SNAP25 antibody (ab5666)
    Flow Cytometry - Anti-SNAP25 antibody (ab5666)

    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.

  • Immunocytochemistry - Anti-SNAP25 antibody (ab5666)
    Immunocytochemistry - Anti-SNAP25 antibody (ab5666)
    Immunlocalization of SNAP-25 in cultured rat hippocampal cells using ab5666.
  • Western blot - Anti-SNAP25 antibody (ab5666)
    Western blot - Anti-SNAP25 antibody (ab5666)This image is courtesy of an anonymous Abreview
    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

    See Abreview

参考文献 (33)

ab5666 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab5666 は 33 報の論文で使用されています。

  • Lauretti E  et al. Extra virgin olive oil improves synaptic activity, short-term plasticity, memory, and neuropathology in a tauopathy model. Aging Cell 19:e13076 (2020). PubMed: 31762202
  • Stévenin V  et al. Dynamic Growth and Shrinkage of the Salmonella-Containing Vacuole Determines the Intracellular Pathogen Niche. Cell Rep 29:3958-3973.e7 (2019). PubMed: 31851926
  • Sifuentes-Dominguez LF  et al. SCGN deficiency results in colitis susceptibility. Elife 8:N/A (2019). PubMed: 31663849
  • Zhou R  et al. Baicalin regulates the dopamine system to control the core symptoms of ADHD. Mol Brain 12:11 (2019). PubMed: 30736828
  • Lin L  et al. Synaptic structure and alterations in the hippocampus in neonatal rats exposed to lipopolysaccharide. Neurosci Lett 709:134364 (2019). PubMed: 31288048
View all Publications for this product

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Western blot abreview for Anti-SNAP25 antibody

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

投稿 Feb 09 2015

Western blot abreview for Anti-SNAP25 antibody

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

投稿 Jun 06 2014

Question

Please provide concentration of ab5666 lot number GR47976-2

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Answer

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 not hesitate to contact us.

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Western blot abreview for Anti-SNAP25 antibody

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Aug 20 2008

Western blot abreview for Anti-SNAP25 antibody

 Excellent
Abreviews
abreview image
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%
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Abcam user community

Verified customer

投稿 Nov 17 2006

Question

BATCH NUMBER 137071 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM no signal by Western blot of mouse trigeminal ganglia protein extracts or immunohistochemistry on mouse brain, using recommended primary antibody dilutions SAMPLE Western blot: mouse trigeminal ganglia total protein extract PRIMARY ANTIBODY SNAP 25 (1:2000) in 1% albumin in 1xPBS containing 0.005% Tween-20, incubation: 2 hours, wash with 1xPBS containing 0.05% Tween-20 DETECTION METHOD Pierce SuperSignal West Dura Extended duration substrate POSITIVE AND NEGATIVE CONTROLS USED I am writing to request information about an appropriate positive control. ANTIBODY STORAGE CONDITIONS 20 ul aliquots stored at -20 C SAMPLE PREPARATION lysis buffer: 1% triton x-100, 2 mM EDTA in 1xPBS Calbiochem protease inhibitor cocktail 3 min boil AMOUNT OF PROTEIN LOADED 100 ug ELECTROPHORESIS/GEL CONDITIONS sample buffer: reducing conditions, specifically, contains Tris-HCl, SDS, glycerol, and B-mercaptoethanol gel: 18% Tris-HCl TRANSFER AND BLOCKING CONDITIONS blocking: 1% albumin in 1xPBS containing 0.005% Tween-20 SECONDARY ANTIBODY Pierce Goat anti-Rabbit HRP-conjugated (1:1500) x 2 hour wash with 1xPBS containing 0.005% Tween-20 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? longer antibody incubation time

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Answer

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 application. We have tested the antibody in WB on rat and mouse brain lysate and therefore with the following recommendations I hope this will improve you signal: - use a stronger lysis buffer to extract the protein (e.g RIPA buffer which contains a strong detergent) -incubate the membrane in blocking buffer (5%BSA) in TBST (Tris buffer with Tween 0.1%) for 1 hr then -incubate the membrane in antibody at 1:1000 overnight (diluted in TBST Tween 0.1%) at 4C Please rinse in TBST too and check that your secondary antibody works well with other primary antibodies. These steps will promote the antibody binding and enhance the signal. If you would like me to look at your IHC protocol I enclose a link to the IHC protocol questionnaire, if you put it to my attention I will look at this as soon as I receive it: https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=5666&mode=questionaire Please let me know if you need further assistance,

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