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RecombinantRabMAb

Recombinant Anti-Otx2 antibody [EPR20375] (ab183951)

Reviews (1) Submit a question References (2)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Microtomy (Formalin-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Western blot - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Immunoprecipitation - Anti-Otx2 antibody [EPR20375] (ab183951)
  • Anti-Otx2 antibody [EPR20375] (ab183951)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20375] to Otx2
  • Suitable for: WB, IHC-P, IP, Microtomy (Formalin-fixed paraffin-embedded sections)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Otx2 antibody [EPR20375]
    See all Otx2 primary antibodies

  • Description

    Rabbit monoclonal [EPR20375] to Otx2

  • Host species

    Rabbit

  • Specificity

    This antibody recognizes isoform 1 and isoform 2 of Otx2.

  • Tested applications

    Suitable for: WB, IHC-P, IP, Microtomy (Formalin-fixed paraffin-embedded sections)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Y79 whole cell lysate. IHC-P: Human retina and retinoblastoma tissues; Mouse E14.5 choroid plexus and retina tissues; Rat E14.5 choroid plexus tissue. IP: Y79 whole cell lysate. FFPE: Mouse e14 whole foetus

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab183951 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
1/5000. Detects a band of approximately 32, 34 kDa (predicted molecular weight: 32 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Microtomy (Formalin-fixed paraffin-embedded sections)
Use at an assay dependent concentration.
Notes
WB
1/5000. Detects a band of approximately 32, 34 kDa (predicted molecular weight: 32 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Microtomy (Formalin-fixed paraffin-embedded sections)
Use at an assay dependent concentration.

Target

  • Function

    Probably plays a role in the development of the brain and the sense organs. Can bind to the BCD target sequence (BTS): 5'-TCTAATCCC-3'.

  • Tissue specificity

    Expressed in brain.

  • Involvement in disease

    Defects in OTX2 are the cause of microphthalmia syndromic type 5 (MCOPS5) [MIM:610125]. Microphthalmia is a clinically heterogeneous disorder of eye formation, ranging from small size of a single eye to complete bilateral absence of ocular tissues. Up to 80% of cases of microphthalia occur in association with syndromes that include non-ocular abnormalities. MCOPS5 patients manifest unilateral or bilateral microphthalmia/clinical anophthalmia and variable additional features including coloboma, microcornea, cataract, retinal dystrophy, hypoplasia or agenesis of the optic nerve, agenesis of the corpus callosum, developmental delay, joint laxity, hypotonia, and seizures.

  • Sequence similarities

    Belongs to the paired homeobox family. Bicoid subfamily.
    Contains 1 homeobox DNA-binding domain.

  • Developmental stage

    Embryo.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CPHD6 antibody
    • Homeobox protein OTX2 antibody
    • MCOPS 5 antibody
    • MCOPS5 antibody
    • MGC45000 antibody
    • Orthodenticle 2 antibody
    • Orthodenticle homeobox 2 antibody
    • Orthodenticle homolog 2 (Drosophila) antibody
    • Orthodenticle homolog 2 antibody
    • Orthodenticle2 antibody
    • Otx 2 antibody
    • otx2 antibody
    • OTX2_HUMAN antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    IHC image of Otx2 staining in a section of formalin-fixed paraffin-embedded normal mouse e14 foetal brain performed on a Leica BOND™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab183951, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Microtomy (Formalin-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Microtomy (Formalin-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    IHC image of Otx2 staining in a section of formalin-fixed paraffin-embedded normal mouse e14 foetus performed on a Leica BOND™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab183951, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    Immunohistochemical analysis of paraffin-embedded rat E14.5 choroid plexus tissue labeling Otx2 with ab183951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on choroid plexus of E14.5 rat is observed [PMID: 23364326]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    Immunohistochemical analysis of paraffin-embedded human retina tissue labeling Otx2 with ab183951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on inner nuclear layer and outer nuclear layer of the human retina is observed [PMID: 19686387]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    Immunohistochemical analysis of paraffin-embedded human retinoblastoma tissue labeling Otx2 with ab183951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on tumor cells of human retinoblastoma is observed [PMID: 19686387]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    Immunohistochemical analysis of paraffin-embedded mouse E14.5 choroid plexus tissue labeling Otx2 with ab183951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on choroid plexus of E14.5 mouse is observed [PMID: 23364326]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody [EPR20375] (ab183951)

    Immunohistochemical analysis of paraffin-embedded mouse retina tissue labeling Otx2 with ab183951 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on nuclear layer of the mouse retina is observed [PMID: 19686387]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Otx2 antibody [EPR20375] (ab183951)
    Western blot - Anti-Otx2 antibody [EPR20375] (ab183951)
    All lanes : Anti-Otx2 antibody [EPR20375] (ab183951) at 1/5000 dilution

    Lane 1 : Y79 (Human retinoblastoma cell line) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 32 kDa
    Observed band size: 32,34 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative control: HeLa (PMID: 12559959).

    This antibody recognizes isoform 1 and isoform 2 of Otx2.

  • Immunoprecipitation - Anti-Otx2 antibody [EPR20375] (ab183951)
    Immunoprecipitation - Anti-Otx2 antibody [EPR20375] (ab183951)

    Otx2 was immunoprecipitated from 0.35 mg of Y79 (Human retinoblastoma cell line) whole cell lysate with ab183951 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab183951 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Y79 whole cell lysate, 10 μg (Input).

    Lane 2: ab183951 IP in Y79 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183951 in Y79 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • Anti-Otx2 antibody [EPR20375] (ab183951)
    Anti-Otx2 antibody [EPR20375] (ab183951)

References (2)

Publishing research using ab183951? Please let us know so that we can cite the reference in this datasheet.

ab183951 has been referenced in 2 publications.

  • Conedera FM  et al. The TGFβ/Notch axis facilitates Müller cell-to-epithelial transition to ultimately form a chronic glial scar. Mol Neurodegener 16:69 (2021). PubMed: 34593012
  • Grall E  et al. Severe head dysgenesis resulting from imbalance between anterior and posterior ontogenetic programs. Cell Death Dis 10:812 (2019). PubMed: 31649239

Customer reviews and Q&As

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Immunocytochemistry abreview for Anti-Otx2 antibody [EPR20375]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cell (ipsc-derived human midbrain dopaminergic neurons)
Permeabilization
Yes - 0.2% triton x-100
Specification
ipsc-derived human midbrain dopaminergic neurons
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Idan Rosh

Verified customer

Submitted Mar 09 2023

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