Overview

  • Product name

    Anti-Zic2 antibody [EPR7790]
    See all Zic2 primary antibodies
  • Description

    Rabbit monoclonal [EPR7790] to Zic2
  • Host species

    Rabbit
  • Specificity

    This antibody detects a 70kDa extra band in some cell lines.
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Zic2 aa 100-200. The exact sequence is proprietary.

  • Positive control

    • SH-SY5Y, C6' Neuro-2a, PANC-1, SK-OV-3 and U-87 MG; Human fetal brain, Human cerebellum and Mouse embryo tissue lysate
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab150404 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 1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 55 kDa).

This antibody detects a 70kDa extra band in some cell lines.

ICC/IF 1/100.

For unpurified use at 1/500 dilution. 

Flow Cyt 1/600.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/1000. 

 

  • Application notes
    Is unsuitable for IHC-P.
  • 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

    • All lanes : Anti-Zic2 antibody [EPR7790] (ab150404) at 1/2000 dilution

      Lane 1 : Human fetal brain tissue lysate
      Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
      Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysate
      Lane 4 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

      Predicted band size: 55 kDa
      Observed band size: 56 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Diluting/Blocking buffer and concentration: 5% NFDM/TBST

      It detects an 70kDa extra band in some cell lines.

       

    • Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Zic2 with Purified ab150404 at 1:100 (3.0 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • All lanes : Anti-Zic2 antibody [EPR7790] (ab150404) at 1/500 dilution

      Lane 1 : Human fetal brain tissue lysate
      Lane 2 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate
      Lane 3 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate
      Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
      Lane 5 : Mouse embryo tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

      Predicted band size: 55 kDa
      Observed band size: 56 kDa why is the actual band size different from the predicted?



      Exposure time:

      Lane1: 3.25 seconds
      Lane2: 20 seconds
      Lane3: 5.5 seconds
      Lane4-5: 60 seconds

      Diluting/Blocking buffer and concentration: 5% NFDM/TBST

      ab150404 detects an 70kDa extra band in some cell lines. The expression profile observed in U-87 MG is consistent withthe literature (PMID: 11038359). Negative control: U-87 MG (PMID: 11038359)

    • Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Zic2 with purified ab150404 at 1:600 dilution (1 µg/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Anti-Zic2 antibody [EPR7790] (ab150404) at 1/15000 dilution (Purified) + Human fetal brain lysates at 15 µg with 5% NFDM/TBST

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 55 kDa
      Observed band size: 63 kDa why is the actual band size different from the predicted?

    • Anti-Zic2 antibody [EPR7790] (ab150404) at 1/20000 dilution (Purified) + Rat brain lysates at 15 µg with 5% NFDM/TBST

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 55 kDa
      Observed band size: 63 kDa why is the actual band size different from the predicted?

    • Anti-Zic2 antibody [EPR7790] (ab150404) at 1/5000 dilution (Purified) + Mouse brain lysates at 15 µg with 5% NFDM/TBST

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 55 kDa
      Observed band size: 63 kDa why is the actual band size different from the predicted?

    • Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (human neuroblastoma) labelling Zic2 with purified ab150404 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

      Control: PBS only

    • Overlay histogram showing SH-SY5Y cells stained with unpurified ab150404 (red line). The cells were fixed with 80% methanol (5 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 ( unpurified ab150404, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    • All lanes : Anti-Zic2 antibody [EPR7790] (ab150404) at 1/1000 dilution (unpurified)

      Lane 1 : Human fetal brain lysate
      Lane 2 : Human cerebellum lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 55 kDa
      Observed band size: ~63 kDa why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Lyu Y  et al. ZIC2 is Essential for Maintenance of Latency and is a Target of an Immediate-Early Protein during KSHV Lytic Reactivation. J Virol N/A:N/A (2017). Read more (PubMed: 28835494) »
    • Ma G  et al. Roles of ZIC family genes in human gastric cancer. Int J Mol Med 38:259-66 (2016). WB ; Human . Read more (PubMed: 27177248) »
    See all 3 Publications for this product

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