Overview

  • Product nameAnti-Sec8 antibody [14G1]
    See all Sec8 primary antibodies
  • Description
    Mouse monoclonal [14G1] to Sec8
  • Tested applicationsSuitable for: IP, WB, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Sheep, Chicken, Cow, Dog, Xenopus laevis, Rainbow Trout
  • Immunogen

    Fusion protein (Recombinant rat rSec8 (his-tagged) expressed in E. coli).

  • Positive control
    • In Western Blot, this antibody gave a positive signal in the following tissue lysates: human brain; human kidney; human placenta; mouse brain.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPreservative: None
    Constituents: 50% Glycerol, PBS, 0.1mM PMSF. pH 7.2
  • Concentration information loading...
  • PurityProtein G purified
  • Purification notesThis antibody has been affinity purified.
  • ClonalityMonoclonal
  • Clone number14G1
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab13254 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 12.5 µg/ml.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 117 kDa (predicted molecular weight: 117 kDa).
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr Use a concentration of 5 µg/ml.

Target

Anti-Sec8 antibody [14G1] images

  • All lanes : Anti-Sec8 antibody [14G1] (ab13254) at 5 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human kidney tissue lysate - total protein (ab30203)
    Lane 3 : Human placenta tissue lysate - total protein (ab29745)
    Lane 4 : Brain (Mouse) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 117 kDa
    Observed band size : 117 kDa
    Additional bands at : 45 kDa,52 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
  • Overlay histogram showing HeLa cells stained with ab13254 (red line). The cells were fixed with 80% methanol (5 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 (ab13254, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunofluorescence analysis of HMVECs, staining Sec8 with ab13254.

    Cells were fixed in paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with blocking buffer. Samples were incubated with primary antibody (1/50) overnight at 4°C. An AlexFluor®-conjugated anti-mouse IgG (1/500) was used as a secondary antibody.
  • Sec8 was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Mouse monoclonal to Sec8 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 ab13254.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
    Band: 117kDa, non specific bands - 45kDa: We are unsure as to the identity of this extra band; Sec8

References for Anti-Sec8 antibody [14G1] (ab13254)

This product has been referenced in:
  • Winkelmann A  et al. Changes in neural network homeostasis trigger neuropsychiatric symptoms. J Clin Invest 124:696-711 (2014). WB ; Mouse . Read more (PubMed: 24430185) »
  • Barkefors I  et al. Exocyst Complex Component 3-like 2 (EXOC3L2) Associates with the Exocyst Complex and Mediates Directional Migration of Endothelial Cells. J Biol Chem 286:24189-99 (2011). ICC/IF, IP ; Mouse . Read more (PubMed: 21566143) »

See all 3 Publications for this product

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