RabMAb

Anti-alpha + beta Synuclein抗体[EP1646Y] (ab51252)

概述

  • 产品名称Anti-alpha + beta Synuclein抗体[EP1646Y]
    参阅全部 alpha + beta Synuclein 一抗
  • 描述
    兔单克隆抗体[EP1646Y] to alpha + beta Synuclein
  • 经测试应用适用于: Flow Cyt, WB, ICC/IFmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human alpha + beta Synuclein aa 1-100 (N terminal). The exact sequence is proprietary.

  • 阳性对照
    • WB: Human fetal brain, human cerebellum, mouse brain and rat brain tissue lysates. ICC/IF: PC12 and rat primary hippocampal neuron cells. Flow Cyt: SH-SY5Y and U87-MG cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-alpha + beta Synuclein antibody (Alexa Fluor® 488) [EP1646Y] (ab199086)

    Anti-alpha + beta Synuclein antibody (Alexa Fluor® 647) [EP1646Y] (ab199304)

性能

应用

Our Abpromise guarantee covers the use of ab51252 in the following tested applications.

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

应用 Ab评论 说明
Flow Cyt 1/30.

For unpurified use at 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).

For unpurified use at 1/50000.

ICC/IF 1/50 - 1/100.
  • 应用说明Is unsuitable for IP.
  • 靶标

    Anti-alpha + beta Synuclein antibody [EP1646Y] 图像

    • Immunocytochemistry/Immunofluorescence analysis of PC-12 cells labelling alpha + beta Synuclein with purified ab51252 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • Overlay histogram showing U87-MG cells fixed in 4% PFA and stained with purified ab51252 at a dilution of 1 in 30 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
    • All lanes : Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/5000 dilution (purified)

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 18 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM /TBST.

    • Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/5000 dilution (purified) + Human cerebellum tissue lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 18 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM /TBST.

    • Flow Cytometry analysis of U87-MG cells labelling alpha + beta Synuclein with purified ab51252 at 1/30 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/50000 dilution (unpurified) + Human fetal brain tissue lysate at 10 µg

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 14 kDa
      Observed band size : 18 kDa (why is the actual band size different from the predicted?)
    • Immunocytochemistry/Immunofluorescence analysis of rat primary hippocampal neurons, staining alpha + beta Synuclein with unpurified ab51252. Cells were fixed, permeabilized, and blocked with 10% donkey serum at room temperature. Cells were incubated with primary antibody (1/1000) at 4°C for 24 hours. A Cy2-conjugated donkey anti-rabbit IgG was used as the secondary antibody.

    • Overlay histogram showing SH-SY5Y cells stained with unpurified ab51252 (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 (unpurified ab51252, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252)参考文献

    This product has been referenced in:
    • Moscovitz O  et al. The Parkinson's-associated protein DJ-1 regulates the 20S proteasome. Nat Commun 6:6609 (2015). WB . Read more (PubMed: 25833141) »
    • Hall H  et al. Characterization of cognitive deficits in rats overexpressing human alpha-synuclein in the ventral tegmental area and medial septum using recombinant adeno-associated viral vectors. PLoS One 8:e64844 (2013). Read more (PubMed: 23705016) »

    See all 7 Publications for this product

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Sample Human Cell (SH-SY5Y)
    Specification SH-SY5Y
    Permeabilization Yes - 0.5% Tween
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Sep 03 2014

    Thank you for your enquiry and for your patience. I can confirm we have the following alpha Synuclein monoclonal antibody which has an N terminal immunogen. ab51252 Rabbit monoclonal [EP1646Y] to alpha Synuclein Reacts with: Mouse, Rat, Human...

    Read More

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