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    otx2-antibody-epr20375-ab183951.pdf

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RecombinantRabMAb

Recombinant Anti-Otx2 antibody [EPR20375] (ab183951)

  • Datasheet
  • SDS
  • Certificate of Compliance
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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

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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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20375
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Sensory System
    • Visual system
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Lineage Markers
    • Ectoderm
    • Developmental Biology
    • Embryogenesis
    • Axis formation and symmetry
    • Developmental Biology
    • Lineage specification
    • Ectoderm
    • Developmental Biology
    • Organogenesis
    • Nervous system development

Associated products

  • Alternative Versions

    • Anti-Otx2 antibody [EPR20375] - BSA and Azide free (ab222495)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Otx2 protein (ab200294)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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.
  • Target information above from: UniProt accession P32243 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5015 Human
    • Entrez Gene: 18424 Mouse
    • Entrez Gene: 305858 Rat
    • Omim: 600037 Human
    • SwissProt: P32243 Human
    • SwissProt: P80206 Mouse
    • SwissProt: Q64201 Rat
    • Unigene: 288655 Human
    • Unigene: 134516 Mouse
    • Unigene: 35222 Rat
    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)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

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

ab183951 has been referenced in 1 publication.

  • Grall E  et al. Severe head dysgenesis resulting from imbalance between anterior and posterior ontogenetic programs. Cell Death Dis 10:812 (2019). PubMed: 31649239

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