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  1. Link

    products/primary-antibodies/otx1--otx2-antibody-ab21990.pdf

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Anti-Otx1 + Otx2 antibody (ab21990)

  • Datasheet
  • SDS
Reviews (10)Q&A (10)References (70)

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Immunoprecipitation - Anti-Otx1 + Otx2 antibody (ab21990)
  • Western blot - Anti-Otx1 + Otx2 antibody (ab21990)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody (ab21990)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody (ab21990)
  • Western blot - Anti-Otx1 + Otx2 antibody (ab21990)
  • Immunocytochemistry/ Immunofluorescence - 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

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-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
  • Immunogen

    Synthetic peptide corresponding to Human Otx1 + Otx2 aa 250 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    Database link: P32243
    (Peptide available as ab24347, ab24348)

  • 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
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • 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

  • ChIP Related Products

    • ChIP Kit (ab500)
  • 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)

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.
ChIP
Use at an assay dependent concentration. PubMed: 18057103
IHC-P (2)
Use at an assay dependent concentration.
ICC/IF (2)
Use a concentration of 0.8 - 4 µg/ml.
WB (2)
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31.6 kDa).
IP
Use a concentration of 5 µg/ml.
Notes
IHC-Fr
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration. PubMed: 18057103
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 0.8 - 4 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31.6 kDa).
IP
Use a concentration of 5 µg/ml.

Target

  • 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.

  • 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
    • SwissProt: Q91981 Zebrafish
    see all
  • Alternative names

    • FLJ38361 antibody
    • Homeobox protein OTX 1 antibody
    • Homeobox protein OTX1 antibody
    • Homeobox protein OTX2 antibody
    • MCOPS 5 antibody
    • MCOPS5 antibody
    • MGC15736 antibody
    • MGC45000 antibody
    • Orthodenticle 1 antibody
    • Orthodenticle 2 antibody
    • Orthodenticle homeobox 1 antibody
    • Orthodenticle homeobox 2 antibody
    • Orthodenticle homolog 1 antibody
    • Orthodenticle homolog 2 (Drosophila) antibody
    • Orthodenticle homolog 2 antibody
    • Orthodenticle1 antibody
    • Orthodenticle2 antibody
    • Otx 1 antibody
    • Otx 2 antibody
    • otx1 antibody
    • OTX1_HUMAN antibody
    • otx2 antibody
    • OTX2_HUMAN antibody
    see all

Images

  • Immunoprecipitation - Anti-Otx1 + Otx2 antibody (ab21990)
    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.

  • Western blot - Anti-Otx1 + Otx2 antibody (ab21990)
    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)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx1 + Otx2 antibody (ab21990)This image is courtesy of Erin Bassett

    ab21990 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)
    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.

  • Western blot - Anti-Otx1 + Otx2 antibody (ab21990)
    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)
    Immunocytochemistry/ Immunofluorescence - Anti-Otx1 + Otx2 antibody (ab21990)This image is courtesy of L. Vallier, University of Cambridge
    The image shows staining of Otx2 in human embryonic stem cells differentiated into neuroectoderm. As would be expected, staining is nuclear.

Protocols

  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (70)

Publishing research using ab21990? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 20 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Pleurodeles waltl Tissue sections (brain)
Permeabilization
Yes - 0.1%Triton
Specification
brain
Blocking step
fish skin gelatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 26°C
Fixative
Formaldehyde
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 Feb 07 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.1% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
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 Feb 28 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Zebra finch Tissue sections (brain)
Antigen retrieval step
None
Permeabilization
No
Specification
brain
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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 Oct 10 2016

Western blot abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Sample
Mouse Tissue lysate - whole (Liver)
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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.

Abcam user community

Verified customer

Submitted Jun 02 2014

Immunohistochemistry (Frozen sections) abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Average
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Mouse Tissue sections (E8.5 mouse embryo brain section)
Specification
E8.5 mouse embryo brain section
Permeabilization
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
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 Nov 05 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Embryonic Stem Cells)
Specification
Embryonic Stem Cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton-X
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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.

Abcam user community

Verified customer

Submitted Mar 29 2012

Question

I was looking for an antibody to Otx2 that has been used and is rated for CHIP and found Abcam Ab21990. However, when I checked the literature cited that Abcam provides to support its claim that is antibody is indeed CHIP grade and has been used for CHIP (ChIP:
Use at an assay dependent dilution. PubMed: 18057103). I found that
this study, while it is study on Otx2, has not performed any CHIP experiments and has not even used any Abcam Antibody.

Read More

Abcam community

Verified customer

Asked on Dec 14 2012

Answer

Thank you for your enquiry. The location of the data is not obvious. The ab21990 ChIP data in PubMed:18057103 is in the supplemental material for this paper. Here is a link:

http://dev.biologists.org/content/135/2/249/suppl/DC1

Chromatin samples were prepared from heads of uninjected stage 22-24 embryos, and imunoprecipitated with either 5 micrograms of control IgG or anti-Otx2 antibody (Abcam, ab21990).

Read More

Abcam Scientific Support

Answered on Dec 14 2012

Question

I am using this antibody per the recommendation of a colleague, but I am getting very different results using the same protocol and samples. Can I please receive another lot to try?

Read More

Abcam community

Verified customer

Asked on Jun 21 2012

Answer

Thank you for contacting Abcam regarding ab21990.


I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on Jun 21 2012

Question

Recently, I’ve been in contact with a customer who’s had some trouble with AB74768; I’ve attached their completed troubleshooting form to the email.

They’re hoping to get a replacement vial, but of AB21990 instead.

Read More

Abcam community

Verified customer

Asked on May 29 2012

Answer

Thank you for contacting us.
I'm sorry to hear that the customer has trouble with this antibody.

The information that was provided by the customer is very helpful to us and I am happy to arrange for a free of charge replacement with ab21990. It will be added to our next shipment to you.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on May 29 2012

Question

I have only been using ab21990 the order confirmation number is .   Please go ahead and send me ab76748 as a replacement. Thank you for sending time on resolving these issues.  

Read More

Abcam community

Verified customer

Asked on Feb 15 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number . To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on Feb 15 2012

1-10 of 20 Abreviews or Q&A

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