Anti-Otx1 + Otx2 抗体 - ChIP Grade (ab21990)

製品の概要

  • 製品名Anti-Otx1 + Otx2 antibody - ChIP Grade
    Otx1 + Otx2 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Otx1 + Otx2 - ChIP Grade
  • 特異性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.
  • アプリケーション適用あり: IHC-Fr, ICC/IF, ChIP, WB, IHC-P, IPmore details
  • 種交差性
    交差種: Mouse, Chicken, Cow, Human, Xenopus laevis
    交差が予測される動物種: Zebrafish
  • 免疫原

    Synthetic peptide corresponding to Human Otx2 aa 250 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P32243
    (Peptide available as ab24347, ab24348)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: Y79.

      This antibody gave a positive signal in the following cell lines: Human Embryonic Stem cells.

      This antibody gave a positive signal in the following tissues: Formalin Fixed Paraffin Embedded Mouse E6 Embryo Normal; Formalin Fixed Paraffin Embedded Mouse E14.5 Embryo Normal

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
IHC-Fr Use a concentration of 0.8 µg/ml.
ICC/IF Use a concentration of 0.8 - 4 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 18057103
WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 31.6 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 20150232
IP Use a concentration of 5 µg/ml.

ターゲット情報

  • 関連性

    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.

  • 参照データベース
  • 別名
    • 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

Anti-Otx1 + Otx2 antibody - ChIP Grade 画像

  • 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).

  • 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.

  • 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.

  • All lanes : Anti-Otx1 + Otx2 antibody - ChIP Grade (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
    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,isoform,non-specific binding).
  • Anti-Otx1 + Otx2 antibody - ChIP Grade (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.
  • The image shows staining of Otx2 in human embryonic stem cells differentiated into neuroectoderm. As would be expected, staining is nuclear.

Anti-Otx1 + Otx2 antibody - ChIP Grade (ab21990) 使用論文

This product has been referenced in:
  • Sanges D  et al. Reprogramming Müller glia via in vivo cell fusion regenerates murine photoreceptors. J Clin Invest 126:3104-3116 (2016). IHC-P ; Mouse . Read more (PubMed: 27427986) »
  • Nagel S  et al. Aberrantly Expressed OTX Homeobox Genes Deregulate B-Cell Differentiation in Hodgkin Lymphoma. PLoS One 10:e0138416 (2015). WB ; Human . Read more (PubMed: 26406991) »

See all 28 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

投稿 Oct 10 2016

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
Username

Abcam user community

Verified customer

投稿 Jun 02 2014

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
Username

Abcam user community

Verified customer

投稿 Nov 05 2013

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

Ch...

Read More

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 th...

Read More

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 ab21...

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

Abcam user community

Verified customer

投稿 Mar 29 2012

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 cau...

Read More

Thank you for getting back to me with these questions.

I have been under the assumption that you havwe been using ab21190 for your experiments, have you been using ab92515 as well?Based on sequence homology ab92515 will recognize both protei...

Read More

Thank you for contacting us.

The immunogen for ab92515 shares 100% identity with OTX1, so cross reactivity is predicted.


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

Read More

1-10 of 18 Abreviews or Q&A

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