Overview

  • Product name
    Anti-14-3-3 Tau antibody [3B9]
    See all 14-3-3 Tau primary antibodies
  • Description
    Mouse monoclonal [3B9] to 14-3-3 Tau
  • Specificity
    ab10439 recognises theta/tau isoform. The antibody does not react with the 14-3-3 zeta isoform.
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Recombinant full length protein corresponding to 14-3-3 Tau.

  • Positive control
    • WB: MBA MD 231, HeLa, A431, Jurkat, Y79 and NIH3T3 whole cell lysates. ICC/IF: HeLa cells. Flow Cyt: SH-SY5Y cells. IP: HeLa cell lysate.
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. 

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. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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

  • Function
    Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathways. Binds to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner. Negatively regulates the kinase activity of PDPK1.
  • Tissue specificity
    Abundantly expressed in brain, heart and pancreas, and at lower levels in kidney and placenta. Up-regulated in the lumbar spinal cord from patients with sporadic amyotrophic lateral sclerosis (ALS) compared with controls, with highest levels of expression in individuals with predominant lower motor neuron involvement.
  • Sequence similarities
    Belongs to the 14-3-3 family.
  • Post-translational
    modifications
    Ser-232 is probably phosphorylated by CK1.
  • Cellular localization
    Cytoplasm. In neurons, axonally transported to the nerve terminals.
  • Information by UniProt
  • Database links
  • Alternative names
    • 14 3 3 protein T cell antibody
    • 14 3 3 protein tau antibody
    • 14 3 3 protein theta antibody
    • 14 3 3 tau antibody
    • 14 3 3 theta antibody
    • 14-3-3 protein T-cell antibody
    • 14-3-3 protein tau antibody
    • 14-3-3 protein theta antibody
    • 1433T_HUMAN antibody
    • 1C5 antibody
    • HS1 antibody
    • KCIP1 antibody
    • Protein HS1 antibody
    • Protein kinase C inhibitor protein 1 antibody
    • Protein tau antibody
    • tyr3/trp5 monooxygenase activation protein, theta antibody
    • Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, theta polypeptide antibody
    • Ywhaq antibody
    see all

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

This product has been referenced in:
  • McFerrin MB  et al. Dysregulation of 14-3-3 proteins in neurodegenerative diseases with Lewy body or Alzheimer pathology. Ann Clin Transl Neurol 4:466-477 (2017). Read more (PubMed: 28695147) »
  • 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 7 Publications for this product

Customer reviews and Q&As

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
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Submitted Dec 12 2014

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