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Invitrogen

OTX2 Monoclonal Antibody (1H12C4B5)

Advanced Verification

3 Published Figures

View all (24) OTX2 antibodies

Questions & Answers

Questions & Answers

Application

OTX2 Antibody in Immunocytochemistry (ICC/IF)

FIGURE: 1 / 10

OTX2 Antibody (MA5-15854) in ICC/IF

Immunofluorescence analysis of HepG2 cells using OTX2 monoclonal antibody (Product # MA5-15854) (Green). Red: actin filaments have been labeled with phalloidin.

OTX2 Antibody in Immunocytochemistry (ICC/IF)

OTX2 Antibody in Immunohistochemistry (Paraffin) (IHC (P))

OTX2 Antibody in Western Blot (WB)

OTX2 Antibody in Western Blot (WB)

OTX2 Antibody in Western Blot (WB)

OTX2 Antibody in Flow Cytometry (Flow)

OTX2 Antibody in Immunohistochemistry (IHC)

OTX2 Antibody in Immunohistochemistry (IHC)

OTX2 Antibody in Immunohistochemistry (IHC)

Product Image Gallery

OTX2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of HepG2 cells using OTX2 monoclonal antibody (Product # MA5-15854) (Green). Red: actin filaments have been labeled with phalloidin.

OTX2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of paraffin-embedded prostate tissues (left) and colon cancer tissues (right) using OTX2 monoclonal antibody (Product # MA5-15854) followed with DAB staining.

OTX2 Antibody in Western Blot (WB) — Western blot was performed using Anti-OTX2 Monoclonal Antibody (1H12C4B5) (Product # MA5-15854) and a 32 kDa band corresponding to OTX2 was observed in mouse retina and rat retina as compared to other tissues. A band at ~25kDa corresponding to circulating tissue IgG was observed in the Mouse tissue lysates. Tissue extracts (30 µg lysate) of Mouse retina (Lane 1), Mouse lung (Lane 2), Mouse eye (Lane 3) Mouse brain (Lane 4) and Rat retina (Lane 5) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution)using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

OTX2 Antibody in Western Blot (WB) — Western blot analysis of OTX2 using OTX2 monoclonal antibody (Product # MA5-15854) in HepG2 (1), Jurkat (2), and NTERA-2 (3) cell lysate.

OTX2 Antibody in Western Blot (WB) — Western blot analysis of OTX2 using a OTX2 monoclonal antibody (Product # MA5-15854) against a human OTX2 (AA: 40-297) recombinant protein.

OTX2 Antibody in Flow Cytometry (Flow) — Flow cytometric analysis of HepG2 cells using OTX2 monoclonal antibody (Product # MA5-15854) (green) and negative control (purple).

OTX2 Antibody in Immunohistochemistry (IHC) — Immunofluorescent analysis of cerebral organoids (A) 26 days in vitro (DIV) COs have ventricular and subventricular zones immunopositive for differentiated neurons (TUJ1 + cells), neural stem cells (SOX2 + cells; green), immature neurons (NeuroD1 + cells; red), proliferative (Ki67 + cells; green), and apoptotic (cleaved caspase 3 + cells; red) cells. An elevated number of cleaved caspase-3+ cells may indicate increased neurosis in larger hCOs. (B) 45DIV COs comprise EAAT1 + and GFAP + astrocytes, as well as late immature neuron markers [NeuN (green) and MAP2 (red)]. (C) COs generated following this protocol are immunopositive for forebrain marker PAX6 and forebrain/midbrain marker OTX2, but not midbrain marker EN1 or hindbrain (GBX2) markers. All scale bars are 100 mum.

OTX2 Antibody in Immunohistochemistry (IHC) — Fig. 10 High activation of the canonical Wnt pathway in hESC-derived organoids causes suppression of ChPe markers. a A schematic presentation of the protocol for the treatment of organoids with the canonical Wnt agonist CHIR. b , c The ChP marker TTR is present in the periphery of a day 25 organoid exposed to 3 muM, but not when exposed to 12 muM CHIR, from day 18. c N = 4 organoids (3 muM CHIR) and 5 organoids (12 muM CHIR treated); were used for immunohistochemistry analysis. d High magnification images (boxed region shown in b) of serial sections of a day 40 organoid exposed to 3 muM reveal the presence of TTR, AQP1, and OTX2 in the periphery. This labeling is reduced or undetectable upon exposure to higher concentrations of CHIR (6/9/12 muM). d N = 14 organoids (3 muM CHIR), 9 organoids (6 muM CHIR), 5 organoids (9 muM CHIR treated), and 14 organoids (12 muM CHIR treated) from 2 independent experiments were used for analysis (each organoid was considered as an independent biological replicate). Additional examples are provided in Supplementary Fig. S10 . e q-PCR analysis shows that the expression of the ChP markers TTR, AQP1 , and OTX2 is upregulated in organoids at day 30 organoids exposed to 3 muM CHIR from day 18. This increased expression is lost upon treatment with 12 muM CHIR for the same period. ( f ) LEF1 expression displays a stepwise increase upon 3 muM and 12 muM CHIR treatment, consistent with increased activation of the canonical Wnt pathway. Bar graphs ( e ,

OTX2 Antibody in Immunohistochemistry (IHC) — Fig. 6 Morphological, immunohistochemical, and molecular features of Rax - creER T2 ::Ctnnb1(ex3) fl/+ SmoM2 fl/+ ( RBS ) ocular lesions match with IO-MEPL as opposed to retinoblastoma. a Morphology of murine RBS lesion after tamoxifen administration on day E8.5 (left column) compared to IO-MEPL (middle column) and human retinoblastoma (right column). Flexner-Wintersteiner-Rosettes in retinoblastoma were smaller in comparison to the pseudostratified rosettes of IO-MEPL and RBS (first row). IO-MEPL and RBS stained strongly for MAP2C, while rosette structures consistently lacked staining signal. In contrast, retinoblastoma displayed strong MAP2C positivity with most prominent staining intensity towards the luminal surfaces of rosettes (second row). SOX2-positive cells were found in IO-MEPL and RBS in a scattered distribution with occasional accentuation of staining intensity in association with rosettes or rosette-like structures, while SOX2 staining was negative in Retinoblastoma (third row). OTX2-positive cells were occasionally found in IO-MEPL in association with rosettes or rosette-like structures and in RBS in the residual retina and NBL-like rosettes. In contrast, retinoblastoma stained negative for OTX2 (fourth row). Scale bar is 100 um. b Hierarchical clustering based on cross-species harmonized gene expression data of this study, GSE30074 [] and GSE172170 []. Ocular lesions of RBS mice formed a distinct cluster with closest similarity to human IO-MEPLs, but not human

OTX2 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissue tested owing to their inherent genetic constitution. Relative expression of OTX2 was observed in Mouse retina in comparison to Mouse Brain, Mouse Eye and Mouse Lung using Anti-OOTX2 Monoclonal Antibody (1H12C4B5) (Product # MA5-15854) in Western Blot. {RE}

Product Details

MA5-15854

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

-

Immunohistochemistry (IHC)

-

View 5 publications 5 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:200-1:1,000

-

Immunocytochemistry (ICC/IF)

1:200-1:1,000

-

Flow Cytometry (Flow)

1:200-1:400

-

ELISA (ELISA)

1:10,000

-

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Mouse

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

1H12C4B5

Immunogen

Purified recombinant fragment of human OTX2 expressed in E. Coli.

Conjugate

Unconjugated

Form

Liquid

Concentration

Conc. not determined

Storage buffer

ascites

Contains

0.03% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Shipping conditions

Ambient (domestic); Wet ice (international)

AB_11155193

Product Specific Information

MA5-15854 targets OTX2 in indirect ELISA, FACS, IF, IHC, and WB applications and shows reactivity with Human samples.

The MA5-15854 immunogen is purified recombinant fragment of human OTX2 expressed in E. Coli. .

MA5-15854 detects OTX2 which has a predicted molecular weight of approximately 32kDa.

Target Information

OTX2 is a 289 amino acid containing protein encoding a member of the bicoid sub-family of homeodomain-containing transcription factors. It has two known alternative splice variants with distinct isoforms though presence of other isoforms cannot be excluded. The protein is an early marker of endodermal differentiation of embryonic stem cells and may play a role in the development of the brain and the sensory organs. The protein may bind to BCD target sequence (BTS): 5-'TCTAATCCC-3'. Defects in OTX2 are the cause of microphthalmia syndromic type 5 (MCOPS5).

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Generation of human induced pluripotent stem cell-derived cerebral organoids for cellular and molecular characterization.

STAR protocols

Anastasaki C,Wilson AF,Chen AS,Wegscheid ML,Gutmann DH

Published figure using OTX2 monoclonal antibody (Product # MA5-15854) in Immunohistochemistry

Fri Mar 18 00:00:00 UTC 2022

Generation of human induced pluripotent stem cell-derived cerebral organoids for cellular and molecular characterization.

STAR protocols

Anastasaki C,Wilson AF,Chen AS,Wegscheid ML,Gutmann DH

Published figure using OTX2 monoclonal antibody (Product # MA5-15854) in Immunohistochemistry

Fri Mar 18 00:00:00 UTC 2022

Constitutive activation of canonical Wnt signaling disrupts choroid plexus epithelial fate.

Nature communications

Parichha A,Suresh V,Chatterjee M,Kshirsagar A,Ben-Reuven L,Olender T,Taketo MM,Radosevic V,Bobic-Rasonja M,Trnski S,Holtzman MJ,Jovanov-Milosevic N,Reiner O,Tole S

Published figure using OTX2 monoclonal antibody (Product # MA5-15854) in Immunohistochemistry

Wed Feb 02 00:00:00 UTC 2022

Constitutive activation of canonical Wnt signaling disrupts choroid plexus epithelial fate.

Nature communications

Parichha A,Suresh V,Chatterjee M,Kshirsagar A,Ben-Reuven L,Olender T,Taketo MM,Radosevic V,Bobic-Rasonja M,Trnski S,Holtzman MJ,Jovanov-Milosevic N,Reiner O,Tole S

Published figure using OTX2 monoclonal antibody (Product # MA5-15854) in Immunohistochemistry

Wed Feb 02 00:00:00 UTC 2022

Co-activation of Sonic hedgehog and Wnt signaling in murine retinal precursor cells drives ocular lesions with features of intraocular medulloepithelioma.

Oncogenesis

Dottermusch M,Sumisławski P,Krevet J,Middelkamp M,Voß H,Siebels B,Bartsch H,Sotlar K,Meyer P,Frank S,Korshunov A,Glatzel M,Schüller U,Neumann JE

Published figure using OTX2 monoclonal antibody (Product # MA5-15854) in Immunohistochemistry

Tue Nov 16 00:00:00 UTC 2021

Patient-derived iPSC-cerebral organoid modeling of the 17q11.2 microdeletion syndrome establishes CRLF3 as a critical regulator of neurogenesis.

Cell reports

Wegscheid ML,Anastasaki C,Hartigan KA,Cobb OM,Papke JB,Traber JN,Morris SM,Gutmann DH

MA5-15854 was used in Immunohistochemistry to identify both neural stem cell (NSC) proliferation and neuronal maturation abnormalities in NF1-TGD hCOs.

Tue Jul 06 00:00:00 UTC 2021

Retinal inputs signal astrocytes to recruit interneurons into visual thalamus.

Proceedings of the National Academy of Sciences of the United States of America

Su J,Charalambakis NE,Sabbagh U,Somaiya RD,Monavarfeshani A,Guido W,Fox MA

MA5-15854 was used in Immunohistochemistry to show that retinal inputs signal through astrocytes to direct the long-range recruitment of interneurons into the visual thalamus.

Tue Feb 04 00:00:00 UTC 2020

Bioinformatics

Protein Aliases: homeobox Otx2; Homeobox protein OTX2; Orthodenticle homolog 2; OTX

Gene Aliases: CPHD6; E130306E05Rik; MCOPS5; OTX2

UniProt ID: (Human) P32243

Entrez Gene ID: (Human) 5015, (Rat) 305858, (Mouse) 18424

Function(s)

RNA polymerase II core promoter proximal region sequence-specific DNA binding transcriptional activator activity, RNA polymerase II core promoter proximal region sequence-specific binding protein binding eukaryotic initiation factor 4E binding DNA binding transcription factor activity, sequence-specific DNA binding sequence-specific DNA binding protein heterodimerization activity homeodomain transcription factor

Process(es)

transcription from RNA polymerase II promoter protein complex assembly axon guidance regulation of smoothened signaling pathway forebrain development midbrain development positive regulation of embryonic development regulation of fibroblast growth factor receptor signaling pathway positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter dopaminergic neuron differentiation primitive streak formation positive regulation of gastrulation regulation of transcription, DNA-templated multicellular organism development cell fate specification regulation of transcription from RNA polymerase II promoter central nervous system development endoderm development organ morphogenesis anterior/posterior pattern specification dorsal/ventral pattern formation metencephalon development cell differentiation neuron differentiation somite rostral/caudal axis specification inner ear morphogenesis eye photoreceptor cell fate commitment cell fate commitment neuron fate commitment neuron fate determination diencephalon morphogenesis anatomical structure development

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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