Anti-Otx1 + Otx2 antibody (ab21990)
Key features and details
- Rabbit polyclonal to Otx1 + Otx2
- Suitable for: IHC-Fr, ChIP, IHC-P, ICC/IF, WB, IP
- Reacts with: Mouse, Chicken, Cow, Human, Xenopus laevis
- Isotype: IgG
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Overview
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Product name
Anti-Otx1 + Otx2 antibody -
Description
Rabbit polyclonal to Otx1 + Otx2 -
Host species
Rabbit -
Specificity
ab21990 is raised against the C-terminal region of the Human Otx2 protein. This product is batch tested in Y79 cell lysate, recombinant Otx1 and overexpressed Otx2. All batches should detect endogenous Otx2, overexpressed Otx2 and recombinant or overexpressed Otx1. -
Tested applications
Suitable for: IHC-Fr, ChIP, IHC-P, ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Mouse, Chicken, Cow, Human, Xenopus laevis
Predicted to work with: Zebrafish
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Immunogen
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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
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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 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
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab21990 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-Fr | (4) |
Use at an assay dependent concentration.
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| ChIP |
Use at an assay dependent concentration. PubMed: 18057103
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| IHC-P | (2) |
Use at an assay dependent concentration.
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| ICC/IF | (2) |
Use a concentration of 0.8 - 4 µg/ml.
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| WB | (2) |
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31.6 kDa).
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| IP |
Use a concentration of 5 µg/ml.
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| Notes |
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IHC-Fr
Use at an assay dependent concentration. |
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ChIP
Use at an assay dependent concentration. PubMed: 18057103 |
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IHC-P
Use at an assay dependent concentration. |
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ICC/IF
Use a concentration of 0.8 - 4 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31.6 kDa). |
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IP
Use a concentration of 5 µg/ml. |
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: 30501 Zebrafish
- 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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Immunoprecipitation - Anti-Otx1 + Otx2 antibody (ab21990)
Otx1 +2 was immunoprecipitated using 0.5mg Y79 whole cell extract, 5µg of Rabbit polyclonal to Otx1 +2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Y79 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab21990.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 37kDa; Otx1 +2, non specific - as present in control (lane 2); 37kDa: We are confident this was due to slight lane contamination and the band seen in the IP lane is our target of interest.
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Western blot - Anti-Otx1 + Otx2 antibody (ab21990)Anti-Otx1 + Otx2 antibody (ab21990) at 1 µg/ml + OTX1 (Human) - 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: 31.6 kDa
Observed band size: 74 kDa why is the actual band size different from the predicted?
This Otx1 recombinant protein has a GST tag causing it to run at ~74kDa. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody (ab21990)This image is courtesy of Erin Bassettab21990 staining Otx2 in embryonic day 14.5 (E14.5) mouse eye by Immunohistochemistry (formalin fixed, paraffin embedded sections).
The embryo was fixed in 10% neutral buffered formalin overnight at room temperature, then paraffin-embedded and sectioned at 4µm. Following deparaffinization and antigen retrieval in a rice steamer in 10mM sodium citrate (pH 6.0), the sections were blocked with 5% normal goat serum for half an hour at room temperature. ab21990 was diluted in PBS at 1/100, and incubated overnight at 4°C. The secondary was goat anti-rabbit Alexa®568, diluted in PBS with 1.5% normal goat serum. The mounting medium contained the nuclear stain DAPI (blue). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody (ab21990)Mouse E6 embryos were sectioned and then fixed in paraformaldehyde. Antigen retrieval was performed using citric acid and the sections permeablized and blocked for 1 hour in serum. Embryos were stained with ab21990 (1/2500) diluted in 10% serum, 1% ovalbumin in PBSTween for 14 hours at 4°C. They were then washed and stained with a goat anti-rabbit biotin conjugated antibody. After using a biotinylated secondary, ABC and then DAB was used for color development.
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Western blot - Anti-Otx1 + Otx2 antibody (ab21990)All lanes : Anti-Otx1 + Otx2 antibody (ab21990) at 1 µg/ml
Lane 1 : Y79 Lysate
Lane 2 : Y79 Lysate with Otx2 peptide (ab24347) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution
Predicted band size: 31.6 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 28 kDa (possible cleavage fragment), 28 kDa (possible isoform), 28 kDa (possible non-specific binding) -
Immunocytochemistry/ Immunofluorescence - Anti-Otx1 + Otx2 antibody (ab21990)This image is courtesy of L. Vallier, University of CambridgeThe image shows staining of Otx2 in human embryonic stem cells differentiated into neuroectoderm. As would be expected, staining is nuclear.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (70)
ab21990 has been referenced in 70 publications.
- Truong V et al. Automating Human Induced Pluripotent Stem Cell Culture and Differentiation of iPSC-Derived Retinal Pigment Epithelium for Personalized Drug Testing. SLAS Technol 26:287-299 (2021). PubMed: 33292045
- Chen X & Emerson MM Notch signaling represses cone photoreceptor formation through the regulation of retinal progenitor cell states. Sci Rep 11:14525 (2021). PubMed: 34267251
- He Q et al. IFI16 promotes human embryonic stem cell trilineage specification through interaction with p53. NPJ Regen Med 5:18 (2020). PubMed: 33298947
- Landowski M et al. Modulation of Tmem135 Leads to Retinal Pigmented Epithelium Pathologies in Mice. Invest Ophthalmol Vis Sci 61:16 (2020). PubMed: 33064130
- Sridhar A et al. Single-Cell Transcriptomic Comparison of Human Fetal Retina, hPSC-Derived Retinal Organoids, and Long-Term Retinal Cultures. Cell Rep 30:1644-1659.e4 (2020). PubMed: 32023475