Overview

  • Product nameAnti-14-3-3 antibody
    See all 14-3-3 primary antibodies
  • Description
    Rabbit polyclonal to 14-3-3
  • SpecificityRecognises all 14-3-3 isoforms.
  • Tested applicationsSuitable for: IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: all Mammals
  • Immunogen

    A mixture of 14-3-3 fusion proteins (beta, eta, tau, and sigma).

Properties

Applications

Our Abpromise guarantee covers the use of ab9063 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 at an assay dependent dilution.
WB 1/1000.
ICC/IF Use a concentration of 5 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. Negative regulator of osteogenesis. Blocks the nuclear translocation of the phosphorylated form (by AKT1) of SRPK2 and antagonizes its stimulatory effect on cyclin D1 expression resulting in blockage of neuronal apoptosis elicited by SRPK2.
  • Sequence similaritiesBelongs to the 14-3-3 family.
  • Post-translational
    modifications
    The alpha, brain-specific form differs from the beta form in being phosphorylated (By similarity). Phosphorylated on Ser-60 by protein kinase C delta type catalytic subunit in a sphingosine-dependent fashion.
    Isoform Short contains a N-acetylmethionine at position 1.
  • Cellular localizationCytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • 14 3 3 antibody
    • 14 3 3 protein beta antibody
    • 14 3 3 protein beta/alpha antibody
    • 14 3 3 protein zeta antibody
    • 14 3 3 zeta antibody
    • 14-3-3 protein beta/alpha antibody
    • 14-3-3 protein/cytosolic phospholipase A2 antibody
    • 1433B_HUMAN antibody
    • GW128 antibody
    • HS1 antibody
    • KCIP 1 antibody
    • KCIP-1 antibody
    • MGC111427 antibody
    • MGC126532 antibody
    • MGC138156 antibody
    • N-terminally processed antibody
    • Protein 1054 antibody
    • Protein kinase C inhibitor protein 1 antibody
    • Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, delta polypeptide antibody
    • Tyrosine 3/tryptophan 5 -monooxygenase activation protein, zeta polypeptide antibody
    • YWHAB antibody
    • YWHAD antibody
    • YWHAZ antibody
    see all

Anti-14-3-3 antibody images

  • All lanes : Anti-14-3-3 antibody (ab9063) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Lung (Human) Tissue Lysate
    Lane 3 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  • 14-3-3 is required for CT-Raf kinase activity. 14-3-3 can be removed from CT-Raf in vitro with the detergent Empigen-BB. Sf9 cells were infected with recombinant baculoviruses encoding either GST or a GST–CT-Raf fusion protein. Forty-eight hours after infection, the GST or GST–CT-Raf proteins were isolated from NP-40 lysates by using glutathione-agarose beads.

    The bead-bound complexes were then washed with either NP-40 lysis buffer (lanes 1 and 2) or NP-40 lysis buffer containing 1% Empigen-BB (lane 3), resolved by SDS-PAGE, transferred to nitrocellulose, and developed with an antibody to either 14-3-3 (upper panel) or Raf (lower panel).

  • ICC/IF image of ab9063 stained PC12 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 (ab9063, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

References for Anti-14-3-3 antibody (ab9063)

This product has been referenced in:
  • Lehmann LH  et al. Essential role of sympathetic endothelin A receptors for adverse cardiac remodeling. Proc Natl Acad Sci U S A 111:13499-504 (2014). IP ; Mouse . Read more (PubMed: 25197047) »
  • Dettmer U  et al. In vivo cross-linking reveals principally oligomeric forms of a-synuclein and ß-synuclein in neurons and non-neural cells. J Biol Chem 288:6371-85 (2013). Read more (PubMed: 23319586) »

See all 11 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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Abcam user community

Verified customer

Submitted Dec 12 2014

Given the similarity of 14-3-3 molecules, it is likely that ab9063 binds to all isoforms and it is also unlikely to find antibodies capable of detecting specifically each isoform. The anti-14-3-3 ab9063 has only been tested against the listed isoforms....

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (skeletal muscle)
Loading amount 50 µg
Specification skeletal muscle
Treatment incubated with +/- insulin and/or +/- crosslinker DSP for 50min
Gel Running Conditions Reduced Denaturing (4-15% precast Tris/HCl gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Feb 06 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Rectal cancer)
Loading amount 20 µg
Specification Rectal cancer
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: ~17°C
Username

Abcam user community

Verified customer

Submitted Jun 14 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Breast cancer)
Loading amount 20 µg
Specification Breast cancer
Gel Running Conditions Non-reduced Denaturing (12 %)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Nov 22 2010

I'm sorry to hear you are having a problem with ab9063 and ab14591, we have not received any complaints about either of those very popular antibodies. Thank you for providing us with your protocol details and with the images of your blots, this is very...

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This antibody is new and we developed it for use in IHC although to date we have not tested it. We hope It will work brilliantly however, given the strong consistent Western results.

All the information available for this product is stated on the datasheet. This antibody was raised against a mixture of beta, eta, tau, and sigma 14-3-3 proteins. Due to the highly conserved nature of mammalian 14-3-3 proteins, it is highly likely th...

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