Anti-RPE65抗体[401.8B11.3D9] (ab13826)

概述

  • 产品名称
    Anti-RPE65抗体[401.8B11.3D9]
    参阅全部 RPE65 一抗
  • 描述
    小鼠单克隆抗体[401.8B11.3D9] to RPE65
  • 宿主
    Mouse
  • 经测试应用
    适用于: ICC/IF, WB, IHC-Fr, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Cow, Human, Pig
    预测可用于: Dog
  • 免疫原

    Tissue, cells or virus corresponding to RPE65. Bovine RPE microsomal membrane proteins.

  • 阳性对照
    • WB: Recombinant human RPE protein expressed in COS-7 cells and bovine RPE membrane fraction. IHC-Fr: Mouse retina tissue. ICC/IF: ARPE19 cells. Flow Cytometry: Mouse retina and choroid cells.
  • 常规说明
    Expression in primary culture of RPE cells is undetectable in Western blot after day 14. Established cell lines are similarly devoid of RPE65 protein, as indicated in western blot. However, mRNA levels are detectable by Northern for at least 7 weeks. (see reference Hamel, C.P., et al. Molecular Cloning and expression of RPE65 a novel retinal pigment epithelium-specific microsomal protein that is post-transcriptionally regulated in vitro. JBC. 268(21): 15751-15757, 1993.)

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. 

性能

应用

Our Abpromise guarantee covers the use of ab13826 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/1000. PubMed: 170320581/1000 (from PubMed:17032058).
WB 1/5000 - 1/10000. Detects a band of approximately 65 kDa.
IHC-Fr 1/250.
Flow Cyt Use at an assay dependent concentration. PubMed: 21494877

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能
    Plays important roles in the production of 11-cis retinal and in visual pigment regeneration. The soluble form binds vitamin A (all-trans-retinol), making it available for LRAT processing to all-trans-retinyl ester. The membrane form, palmitoylated by LRAT, binds all-trans-retinyl esters, making them available for IMH (isomerohydrolase) processing to all-cis-retinol. The soluble form is regenerated by transferring its palmitoyl groups onto 11-cis-retinol, a reaction catalyzed by LRAT. The enzymatic activity is linearly dependent of the expression levels and membrane association.
  • 组织特异性
    Retinal pigment epithelium specific.
  • 疾病相关
    Defects in RPE65 are the cause of Leber congenital amaurosis type 2 (LCA2) [MIM:204100]. LCA designates a clinically and genetically heterogeneous group of childhood retinal degenerations, generally inherited in an autosomal recessive manner. Affected infants have little or no retinal photoreceptor function as tested by electroretinography. LCA represents the most common genetic cause of congenital visual impairment in infants and children.
    Defects in RPE65 are the cause of retinitis pigmentosa type 20 (RP20) [MIM:613794]. RP leads to degeneration of retinal photoreceptor cells. Patients typically have night vision blindness and loss of midperipheral visual field. As their condition progresses, they lose their far peripheral visual field and eventually central vision as well. RP20 inheritance is autosomal dominant.
  • 序列相似性
    Belongs to the carotenoid oxygenase family.
  • 翻译后修饰
    Palmitoylation by LRAT regulates ligand binding specificity; the palmitoylated form (membrane form) specifically binds all-trans-retinyl-palmitate, while the soluble unpalmitoylated form binds all-trans-retinol (vitamin A).
  • 细胞定位
    Cytoplasm. Cell membrane. Attached to the membrane by a lipid anchor when palmitoylated (membrane form), soluble when unpalmitoylated.
  • Information by UniProt
  • 数据库链接
  • 别名
    • All-trans-retinyl-palmitate hydrolase antibody
    • LCA 2 antibody
    • LCA2 antibody
    • Leber congenital amaurosis antibody
    • mRPE 65 antibody
    • mRPE65 antibody
    • p63 antibody
    • rd 12 antibody
    • rd12 antibody
    • Retinal pigment epithelium specific 61 kDa protein antibody
    • Retinal pigment epithelium specific 65 kDa protein antibody
    • Retinal pigment epithelium specific protein antibody
    • Retinal pigment epithelium specific protein 65kDa antibody
    • Retinal pigment epithelium-specific 65 kDa protein antibody
    • Retinitis pigmentosa 20 antibody
    • Retinoid isomerohydrolase antibody
    • Retinol isomerase antibody
    • RP 20 antibody
    • RP20 antibody
    • RPE 65 antibody
    • RPE65 antibody
    • RPE65_HUMAN antibody
    • sRPE 65 antibody
    • sRPE65 antibody
    see all

图片

  • ARPE19 (spontaneously arising human retinal pigment epithelia cell line) cells stained for RPE65 (green) using ab13826 (1/100 dilution) in ICC/IF.

    Cells were fixed in 4% PFA for 10 minutes, permeabilized in PBS containing 0.2% Triton X-100 (PBS-T) for 10 minutes, then blocked for 1 hour in 10% normal goat serum containing 1% BSA in PBS-T. Incubated with primary antibody at 4°C.

  • Frozen sectioned mouse retina tissue stained for RPE65 with ab13826 at a 1/250 dilution in immunohistochemical analysis.

  • Flow Cytometry analysis of mouse retina and choroid cells stained for RPE65 using ab13826.

    RPE65 (right) versus control (left) in the choroid (top) and the retina (bottom).

  • All lanes : Anti-RPE65 antibody [401.8B11.3D9] (ab13826) at 1/5000 dilution

    Lane 1 : Recombinant human RPE-transfected COS-7 (African green monkey kidney fibroblast-like cell line) cell lysate at 20 µg
    Lane 2 : Bovine retinal pigment epithelium membrane fraction at 5 µg

    Secondary
    All lanes : Alkaline phosphatase conjugated goat-anti Mouse IgG

    Observed band size: 60 kDa (why is the actual band size different from the predicted?)



    NBT/BCIP substrate

文献

This product has been referenced in:
  • Kelley RA  et al. DNA nanoparticles are safe and nontoxic in non-human primate eyes. Int J Nanomedicine 13:1361-1379 (2018). Read more (PubMed: 29563793) »
  • Balmer D  et al. Bis-Retinoid A2E Induces an Increase of Basic Fibroblast Growth Factor via Inhibition of Extracellular Signal-Regulated Kinases 1/2 Pathway in Retinal Pigment Epithelium Cells and Facilitates Phagocytosis. Front Aging Neurosci 9:43 (2017). WB ; Mouse . Read more (PubMed: 28298893) »

See all 30 Publications for this product

客户评价及客户问答

Merci de nous avoir contactés.

L'anticorps 13826 est un monoclonal, il ne reconnait donc pas toute la séquence mais uniquement une région très précise de l'immunogène qui a été utilis&eac...

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Merci de votre réponse.

Nous n'avons pas un autre lot de l'ab13826 en stock. Comme vous l'avez souhaité, je vous ai envoyé donc une unité de l'ab78036. Je me permets de vous transférer le numéro de co...

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Merci d'avoir pris le temps de mettre ensemble ce dossier et de nous avoir contactés. Je suis désolée d'apprendre que vous avez eu des difficultés à obtenir de bons résultats avec ab13826.

Aprè...

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Thank you for your inquiry and your patience while we were consulting with the laboratory. I am sorry to hear you are experiencing difficulties with one of our products.

Unfortunately we do not know whether this antibody does react with prote...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (RPE cells extracted from mouse eyes)
Loading amount
25 µg
Specification
RPE cells extracted from mouse eyes
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

提交于 Nov 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mouse eye sections)
Specification
mouse eye sections
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TX100
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

提交于 Nov 25 2010

I'm afraid the epitope recognized by ab13826 has not been mapped yet so we do not know if the antibody is likely to cross react with Xenopus protein. The immunogen used to generate the antibody was whole Bovine RPE microsomal membrane proteins therefor...

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I'm sorry to hear you are having a problem with ab13826 in IHC, we have not tested this antibody in IHC or ICC so it is difficult to find out what might be the problem, the fact that the antibody is a mouse monoclonal and you use it on mouse tissue is ...

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

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