Recombinant Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3347] to Otx1 + Otx2 - ChIP Grade
- Suitable for: WB, IHC-P, IP, ChIP
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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Product name
Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade
See all Otx1 + Otx2 primary antibodies -
Description
Rabbit monoclonal [EPR3347] to Otx1 + Otx2 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, ChIPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
This information is considered to be commercially sensitive.
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Positive control
- WB: Y79 whole cell lysate and Otx1 recombinant protein. IHC-P: Human retina, human retinoblastoma, human cerebral cortex, mouse retina, mouse E14.5 choroid plexus and rat E14.5 choroid plexus. ChIP: Mouse stem cell D3. IP: Y79 whole cell lysate
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3347 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92515 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 |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 32 kDa.
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IHC-P |
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
We recommend 1/500 dilution for human and mouse tissues, and 1/1000 for rat tissues. |
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IP |
1/20.
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ChIP |
Use 5 µg for 25 µg of chromatin.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 32 kDa. |
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. We recommend 1/500 dilution for human and mouse tissues, and 1/1000 for rat tissues. |
IP
1/20. |
ChIP
Use 5 µg for 25 µg of chromatin. |
Target
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Relevance
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.
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.
Similarity: Belongs to the paired homeobox family. Bicoid subfamily. Contains 1 homeobox DNA-binding domain.
Developmental stage: Embryo.
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Database links
- Entrez Gene: 5013 Human
- Entrez Gene: 5015 Human
- Entrez Gene: 18423 Mouse
- Entrez Gene: 25646 Rat
- SwissProt: P32242 Human
- SwissProt: P32243 Human
- SwissProt: P80205 Mouse
- SwissProt: P80206 Mouse
see all -
Alternative names
- FLJ38361 antibody
- Homeobox protein OTX 1 antibody
- Homeobox protein OTX1 antibody
see all
Images
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Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515) at 1/2500 dilution (purified) + Y79 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human retina tissue labelling Otx1 + Otx2 with purified ab92515 at a dilution 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Nuclear staining on inner nuclear layer and outer nuclear layer of the human retina.
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Chromatin was prepared from mouse stem cell D3 according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab92515 (blue), and 20µl of Anti rabbit IgG sepharose beads. 5μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
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Otx1 + Otx2 was immunoprecipitated from Y79(Human retinoblastoma retinoblastoma)whole cell lysate with ab92515 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab92515 at 1/500 dilution. Secondary antibody details (ab131366), was used as secondary antibody at 1/1000 dilution.
Blocking/Diluting Buffer and concentration: 5% NFDM /TBST.
Lanes 1-2 : Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515) at 1/500 dilution (purified)
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab92515
Lane 1 : Y79 (Human retinoblastoma retinoblastoma) whole cell lysate
Lanes 2-3 : Y79 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution -
Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515) at 1 µg/ml (unpurified) + OTX1 Recombinant Protein at 0.1 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human retinoblastoma tissue labelling Otx1 + Otx2 with purified ab92515 at a dilution 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Nuclear staining on tumor cells of human retinoblastoma.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse retina tissue labelling Otx1 + Otx2 with purified ab92515 at a dilution 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Nuclear staining on nuclear layer of the mouse retina.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse E14.5 choroid plexus tissue labelling Otx1 + Otx2 with purified ab92515 at a dilution 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Nuclear staining on mouse E14.5 choroid plexus.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat E14.5 choroid plexus tissue labelling Otx1 + Otx2 with purified ab92515 at a dilution 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Nuclear staining on rat E14.5 choroid plexus.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody [EPR3347] - ChIP Grade (ab92515)Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab92515 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab92515 has been referenced in 1 publication.
- Saini JS et al. Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-Related Macular Degeneration. Cell Stem Cell : (2017). PubMed: 28132833