Overview

  • Product nameAnti-Otx2 antibody
    See all Otx2 primary antibodies
  • Description
    Rabbit polyclonal to Otx2
  • Tested applicationsSuitable 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 conjugated to KLH derived from within residues 200 to the C-terminus of Human Otx2.

  • 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.

Properties

Applications

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 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

  • FunctionProbably 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 specificityExpressed in brain.
  • Involvement in diseaseDefects 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 similaritiesBelongs to the paired homeobox family. Bicoid subfamily.
    Contains 1 homeobox DNA-binding domain.
  • Developmental stageEmbryo.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • 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

Anti-Otx2 antibody images

  • 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.

  • 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).

  • 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).

  • 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
    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.
  • 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
    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.

References for Anti-Otx2 antibody (ab114138)

ab114138 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (E10.5 mouse embryo brain section)
Permeabilization Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Specification E10.5 mouse embryo brain section
Blocking step NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Nov 04 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Mouse Cell (mouse epiblast stem cells)
Specification mouse epiblast stem cells
Permeabilization Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Fixative Paraformaldehyde
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Submitted Nov 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Mr. Carl Hobbs

Verified customer

Submitted Apr 12 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Mr. Carl Hobbs

Verified customer

Submitted Apr 12 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"