Anti-Zic2 antibody (ab10550)

Reviews (1)Q&A (1)References (2)

Overview

  • Product name

    Anti-Zic2 antibody
    See all Zic2 primary antibodies

  • Description

    Rabbit polyclonal to Zic2

  • Host species

    Rabbit

  • Specificity

    Recognises a band at about 60kD in mouse brain lysate (but not in rat brain lysate). A lower band is also detected, the identity of this is unknown.

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Mouse
    Predicted to work with: Cow, Human, Zebrafish

  • Immunogen

    Synthetic peptide:

    GRSEQYRQVASPRTDPYS

    conjugated to KLH, corresponding to amino acids 190-207 of Mouse Zic2.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

Properties

Applications

Our Abpromise guarantee covers the use of ab10550 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
WB Use at an assay dependent dilution. Detects a band of approximately 60 kDa (predicted molecular weight: 55 kDa).

Target

  • Function

    Acts as a transcriptional activator or repressor. Plays important roles in the early stage of organogenesis of the CNS. Activates the transcription of the serotonin transporter SERT in uncrossed ipsilateral retinal ganglion cells (iRGCs) to refine eye-specific projections in primary visual targets. Its transcriptional activity is repressed by MDFIC. Involved in the formation of the ipsilateral retinal projection at the optic chiasm midline. Drives the expression of EPHB1 on ipsilaterally projecting growth cones. Binds to the minimal GLI-consensus sequence 5'-TGGGTGGTC-3'. Associates to the basal SERT promoter region from ventrotemporal retinal segments of retinal embryos.

  • Involvement in disease

    Defects in ZIC2 are a cause of holoprosencephaly type 5 (HPE5) [MIM:609637]. A structural anomaly of the brain, in which the developing forebrain fails to correctly separate into right and left hemispheres. Holoprosencephaly is genetically heterogeneous and associated with several distinct facies and phenotypic variability. Although severe facial anomalies are common in HPE, patients with ZINC2 mutations have relatively normal faces.

  • Sequence similarities

    Belongs to the GLI C2H2-type zinc-finger protein family.
    Contains 5 C2H2-type zinc fingers.

  • Domain

    The C2H2-type 3, 4 and 5 zinc finger domains are necessary for transcription activation.

  • Post-translational
    modifications

    Phosphorylated.
    Ubiquitinated by RNF180, leading to its degradation.

  • Cellular localization

    Nucleus. Cytoplasm. Localizes in the cytoplasm in presence of MDFIC overexpression. Both phosphorylated and unphosphorylated forms are localized in the nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • HPE 5 antibody
      • HPE5 antibody
      • Odd paired homolog Drosophila antibody
      • Zic 2 antibody
      • Zic family member 2 (odd paired Drosophila homolog) antibody
      • Zic family member 2 antibody
      • ZIC2 antibody
      • ZIC2_HUMAN antibody
      • Zinc finger protein of the cerebellum 2 antibody
      • Zinc finger protein ZIC 2 antibody
      • Zinc finger protein Zic2 antibody
      see all

    Images

    • Western blot - Anti-Zic2 antibody (ab10550)
      Western blot - Anti-Zic2 antibody (ab10550)

      Western blot using ab10550 at 1/500.

      Lane 1 : Mouse Brain tissue lysate
      Lane 2 : Rat Brain tissue lysate

      The upper band in the mouse brain lysate represents Zic2.

      Secondary ab: Goat anti-rabbit IgG ab6721 (1/5000)
      Exposure time: 5 minutes
      Expected molecular weight: 55 kDa
      20µg of cell lysate per lane

      Western blot using ab10550 at 1/500.

      Lane 1 : Mouse Brain tissue lysate
      Lane 2 : Rat Brain tissue lysate

      The upper band in the mouse brain lysate represents Zic2.

      Secondary ab: Goat anti-rabbit IgG ab6721 (1/5000)
      Exposure time: 5 minutes
      Expected molecular weight: 55 kDa
      20µg of cell lysate per lane

    References

    This product has been referenced in:

    • Sun Y  et al. Genetically engineered recombinant adenovirus expressing interleukin-2 for hepatocellular carcinoma therapy. Mol Med Rep 17:300-306 (2018). Read more (PubMed: 29115604) »
    • Glendining KA  et al. Downstream mediators of Ten-m3 signalling in the developing visual pathway. BMC Neurosci 18:78 (2017). Read more (PubMed: 29207951) »
    See all 2 Publications for this product

    Publishing research using ab10550? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    Hi,

    I have a customer enquiry about the use of the above AB10550 - https://www.abcam.com/Zic2-antibody-ab10550.html – in IP experiment.

    The customer used this Ab to do pull down from mouse brain lysate, followed by a WB (using same Ab) but could not detect anything in WB. They loaded 5ug original lysate / lane as +ve control and detected an expected 60 kDa band. Nothing in eluate and supernatant fraction of their processed (IP’d) sample – they estimated that the protein load should not have been much less than in their +ve control lane (they started out with 300 ug of lysate.

    A few years ago, they used AB12072 (now discontinued) for the same purpose and that antibody worked for IP. They are confident that their IP system is working since they used it with other proteins (Zic3 etc) and they worked.

    My questions are:

    1. This Ab had not been tested for IP, so can you predict whether it could work?

    2. Re the now discontinued AB12072, had it been tested for IP?

    3. What was the immunogen for AB12072

    If you can throw some light on this issue, that would be good. If you think it is worthwhile to get the customer to fill in a technical questionnaire, I can get them to do that.

    I look forward to hearing your comments.

    Kind regards,

    Read More
    Answer

    Thank you for contacting us.

    As you have noticed, the Anti-Zic2 antibody (ab10550) has as yet not been tested in IP. This does not necessarily mean that it would not work, simply that we have not tested it and would not be able to guarantee its performance.

    It is difficult to predict if the antibody would work in this application. We have only tested it in Western blotting. I would suggest it might be worthwhile for the customer to try performing a Western blot just to make sure that the antibody is active.

    The immunogen used to raise the ab10550 is a synthetic peptide composed of the residues 190-207 of Mouse Zic2 whilst the antibody ab12072 was raised against the peptide taken from residues 162-176 of Human Zic2. Ab12072 had also not been tested for IP but it had been tested in IHC-Fr which indicates that it is likely to be able to recognise the native structure of the protein.

    I hope this information has been of some help. If you have any further questions, please do not hesitate to ask.

    Read More

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Zic2 antibody

     Poor
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mouse forebrain E17)
    Specification
    Mouse forebrain E17
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium Citrate
    Permeabilization
    Yes - 0.025% TX100
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
    Read More

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    Submitted Aug 21 2008

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