Overview

  • Product nameAnti-14-3-3 Tau antibody [3B9]
    See all 14-3-3 Tau primary antibodies
  • Description
    Mouse monoclonal [3B9] to 14-3-3 Tau
  • Specificityab10439 recognises theta/tau isoform. The antibody does not react with the 14-3-3 zeta isoform.
  • Tested applicationsFlow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Recombinant full length protein.

  • Positive control
    • Whole extracts of mouse 3T3, rat PC12, human Jurkat, or normal fibroblasts or bovine MDBK cells. This antibody gave a positive signal during western blot in the following whole cell lysates: MBA MD 231; HeLa; A431; Jurkat; Y79; NIH3T3.
  • General notesAb81216-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab10439 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
Flow Cyt Use 1µg for 106 cells.
IP Use a concentration of 5 µg/ml.
WB 1/5000. Detects a band of approximately 31 kDa (predicted molecular weight: 28 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 19960480
ICC/IF Use a concentration of 1 µg/ml.

Target

Anti-14-3-3 Tau antibody [3B9] images

  • 14-3-3 Tau was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to 14-3-3 Tau 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, Hela whole cell extract 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10439.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 31kDa, non specific band - 26kDa: We are unsure as to the identity of this extra band; 14-3-3 Tau

  • ICC/IF image of ab10439 stained HeLa cells. The cells were 100% methanol fixed (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 (ab10439, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • All lanes : Anti-14-3-3 Tau antibody [3B9] (ab10439)

    Lane 1 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 28 kDa
    Observed band size : 31 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 120 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • Overlay histogram showing SH-SY5Y cells stained with ab10439 (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 (ab10439, 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 SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-14-3-3 Tau antibody [3B9] (ab10439)

This product has been referenced in:
  • Slone SR  et al. 14-3-3theta Protects against Neurotoxicity in a Cellular Parkinson's Disease Model through Inhibition of the Apoptotic Factor Bax. PLoS One 6:e21720 (2011). WB ; Human . Read more (PubMed: 21799745) »
  • Broadbelt KG  et al. Brainstem deficiency of the 14-3-3 regulator of serotonin synthesis: a proteomics analysis in the sudden infant death syndrome. Mol Cell Proteomics : (2011). WB ; Human . Read more (PubMed: 21976671) »

See all 6 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (macrophage)
Specification macrophage
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 12 2014

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