Overview

  • Product name
    Anti-14-3-3 (phospho S58) antibody
    See all 14-3-3 primary antibodies
  • Description
    Rabbit polyclonal to 14-3-3 (phospho S58)
  • Host species
    Rabbit
  • Specificity
    Specific for the ~29k 14-3-3 protein phosphorylated at Ser58. Immunolabeling is blocked by the phosphopeptide used as antigen but not by the corresponding dephosphopeptide.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Sheep, Chicken, Cow, Dog, Xenopus laevis, Non human primates
  • Immunogen

    Phosphopeptide corresponding to amino acid residues surrounding the phosphorylation site Ser58 of rat 14-3-3 protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab30554 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
ICC/IF Use at an assay dependent concentration. PubMed: 21118956
WB 1/1000. Detects a band of approximately 29 kDa.
IHC-Fr 1/500.

Target

Images

  • All lanes : Anti-14-3-3 (phospho S58) antibody (ab30554) at 1/1000 dilution

    Lane 1 : Rat brain stem lysate
    Lane 2 : Rat brain stem lysate with the phosphopeptide used as the antigen

    Observed band size: 29 kDa
    why is the actual band size different from the predicted?

  • ab30554 staining 14-3-3 (phospho S58) in Mouse hippocampal tissue by Immunohistochemistry (Frozen sections). ab30554 is green, red staining is GFAP and nuclei are stained blue with DAPI.
  • ab30554 staining 14-3-3 (phospho S58) in human U2OS cells by Immunocytochemistry/ Immunofluorescence.
    U2OS cells were plated on coverslips and fixed by sequential incubations with 4% formaldehyde in PBS without calcium and magnesium [PBS(-)], 0.1% Triton X-100 in PBS(-) and 0.05% Tween-20 in PBS(-), each for 10 minutes at room temperature. After being washed, cells were incubated with ab30554, followed by incubation with Alexa Fluor 488–conjugated anti-rabbit IgG in TBST (100 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.05% Tween-20) containing 5% FBS.
  • All lanes : Anti-14-3-3 (phospho S58) antibody (ab30554) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate (20ug)
    Lane 3 : Mouse brain homogenate (20ug)

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 28 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute

References

This product has been referenced in:
  • Civiero L  et al. PAK6 Phosphorylates 14-3-3? to Regulate Steady State Phosphorylation of LRRK2. Front Mol Neurosci 10:417 (2017). Read more (PubMed: 29311810) »
  • Okada N  et al. A novel Chk1/2-Lats2-14-3-3 signaling pathway regulates P-body formation in response to UV damage. J Cell Sci 124:57-67 (2011). WB, ICC/IF ; Human . Read more (PubMed: 21118956) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Sample
Mouse Cell (Mouse colon.)
Specification
Mouse colon.
Permeabilization
No
Fixative
PFA/Ethanol

Abcam user community

Verified customer

Submitted Aug 01 2014

Answer

Thank you for your enquiry. I have been in touch with the source of this antibody and I have determined that by immunoblotting this antibody is blocked by the phosphopeptide used as the antigen but NOT by the corresponding dephosphopeptide. We do not offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. We do, however, have our Abreview system, where researchers can submit reviews of our products in exchange for Abpoints. Abpoints can be redeemed for rewards such as Amazon.com gift certificates or discounts on future antibody purchases. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview right from the online datasheet. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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

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