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    otx2-antibody-ab114138.pdf

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Anti-Otx2 antibody (ab114138)

  • Datasheet
  • SDS
Reviews (6) Submit a question References (4)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
  • Western blot - Anti-Otx2 antibody (ab114138)
  • Western blot - Anti-Otx2 antibody (ab114138)

Key features and details

  • Rabbit polyclonal to Otx2
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human, Recombinant fragment
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Otx2 antibody [EPR20375] (ab183951)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Otx2 antibody
    See all Otx2 primary antibodies
  • Description

    Rabbit polyclonal to Otx2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment
    Predicted to work with: Horse, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster, Orangutan
  • Immunogen

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

  • Positive control

    • This antibody gave a positive signal in Otx2 recombinant protein as well as Y79 and WERI whole cell lysates and the following tissue lysates: Mouse Embryonic Brain E10; Mouse Embryonic Brain E14; Mouse Embryonic Brain E16; Mouse Eye. FFPE: Mouse e14 whole foetus
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • 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

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Otx2 protein (ab200294)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab114138 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
IHC-P (3)
1/500 - 1/600. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 32 kDa).
Notes
IHC-P
1/500 - 1/600. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 32 kDa).

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 (ab114138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)

    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 ab114138, 2.5ug/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 (ab114138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)

    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 ab114138, 2.5ug/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 (ab114138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)This image is courtesy of Prof. Antonio Simeone, Institute of Genetics and Biophysics, Naples, Italy.

    IHC-P image of Otx2 antibody (ab114138) on paraffin-embedded mouse embryo sections. The image on the left is of a WT embryo, whilst on the right is an embryo where Otx2 has been replaced with human Otx1 (hOtx1Otx2/Otx2). The sections underwent heat mediated antigen retrieval using Citrate Buffer, primary antibody was used at 1:500 dilution and Biocare Mach4 Kit was used to detect the primary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)This image is courtesy of Carl Hobbs (Kings College London, United Kingdom)

    IHC-P image of Otx2 staining on Rat E12 embryo using ab114138 (1:1500). The sections were deparffinized and subjected to heat mediated antigen retrieval using citric acid. The section were then blocked using 1% BSA for 10 mins at 21°C. ab114138 was diluted 1:1500 using TBS/BSA/Azide and incubated for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to Biotin (1:250).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (ab114138)This image is courtesy of Carl Hobbs (Kings College London, United Kingdom)

    IHC-P image of Otx2 staining on mouse E14 embryo using ab114138 (1:1000). The sections were deparffinized and subjected to heat mediated antigen retrieval using citric acid. The section were then blocked using 1% BSA for 10 mins at 21°C. ab114138 was diluted 1:1000 using TBS/BSA/Azide and  incubated for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to Biotin (1:250).

  • Western blot - Anti-Otx2 antibody (ab114138)
    Western blot - Anti-Otx2 antibody (ab114138)
    All lanes : Anti-Otx2 antibody (ab114138) at 1 µg/ml

    Lane 1 : OTX1 (Human) - Recombinant Protein
    Lane 2 : OTX2 Transient Overexpression Lysate

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 32 kDa
    Observed band size: 37 kDa why is the actual band size different from the predicted?
    Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes


    Lane 1 represents the Otx1 Recombinat protein which was used as a negative control for ab114138. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab114138 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  • Western blot - Anti-Otx2 antibody (ab114138)
    Western blot - Anti-Otx2 antibody (ab114138)
    All lanes : Anti-Otx2 antibody (ab114138) at 1 µg/ml

    Lane 1 : E10 Mouse Embryo Brain Tissue Lysate
    Lane 2 : E14 Mouse Embryo Brain Tissue Lysate
    Lane 3 : E16 Ms Embryo Brain Tissue Lysate
    Lane 4 : Mouse eye tissue lysate - total protein (ab4029)
    Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 6 : WERI (Human Retinoblastoma) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 32 kDa
    Observed band size: 37 kDa why is the actual band size different from the predicted?
    Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab114138 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab114138? Please let us know so that we can cite the reference in this datasheet.

ab114138 has been referenced in 4 publications.

  • Toms D  et al. Automated Hypothesis Generation to Identify Signals Relevant in the Development of Mammalian Cell and Tissue Bioprocesses, With Validation in a Retinal Culture System. Front Bioeng Biotechnol 8:534 (2020). PubMed: 32582664
  • Imbriglio T  et al. Developmental abnormalities in cortical GABAergic system in mice lacking mGlu3 metabotropic glutamate receptors. FASEB J 33:14204-14220 (2019). PubMed: 31665922
  • Zhu Z  et al. Vasopressin regulates hypothalamic GnRH synthesis: Histomorphological evidence in hypothalamus and biological effects in GT1-7 cells. Life Sci 227:166-174 (2019). PubMed: 31026452
  • Navarra A  et al. Hmga2 is necessary for Otx2-dependent exit of embryonic stem cells from the pluripotent ground state. BMC Biol 14:24 (2016). WB . PubMed: 27036552

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Otx2 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (retina)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
Yes - 0.3% triton X-100
Specification
retina
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.

Abcam user community

Verified customer

Submitted Mar 24 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Otx2 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain E12)
Specification
Brain E12
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
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. Carl Hobbs

Verified customer

Submitted Apr 12 2013

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Otx2 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (E14 embryo head)
Specification
E14 embryo head
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
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. Carl Hobbs

Verified customer

Submitted Apr 12 2013

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