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.

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

  • FunctionAdapter 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 specificityAbundantly 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 similaritiesBelongs to the 14-3-3 family.
  • Post-translational
    modifications
    Ser-232 is probably phosphorylated by CK1.
  • Cellular localizationCytoplasm. 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 Binding Motif antibody
    • 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
    • 1433T_HUMAN antibody
    • 1C5 antibody
    • HS1 antibody
    • KCIP1 antibody
    • Protein HS1 antibody
    • Protein kinase C inhibitor protein 1 antibody
    • Protein tau antibody
    • T-CELL antibody
    • tyr3/trp5 monooxygenase activation protein, theta antibody
    • Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, theta polypeptide antibody
    • YWHAQ antibody
    see all

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
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Submitted Dec 12 2014

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