Anti-RPE65抗体[401.8B11.3D9] (ab78036)

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
  • 经测试应用适用于: Flow Cyt, ICC/IF, Other, WB, IP, ELISA, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Cow, Human, Xenopus laevis, Monkey
  • 免疫原

    Bovine RPE microsomal membranes.

  • 阳性对照
    • Eye. Bovine RPE membrane. Recombinant human RPE65 expressed in COS7 cells

性能

应用

Our Abpromise guarantee covers the use of ab78036 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
Flow Cyt Use 2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.
AP Use at an assay dependent concentration.
Other Use at an assay dependent concentration.
WB 1/5000 - 1/20000. Detects a band of approximately 65 kDa (predicted molecular weight: 61 kDa).
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-Fr 1/250 - 1/500.

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

Anti-RPE65 antibody [401.8B11.3D9] 图像

  • All lanes : Anti-RPE65 antibody [401.8B11.3D9] (ab78036) at 1/5000 dilution

    Lane 1 : Bovine RPE membrane at 5 µg
    Lane 2 : Recombinant human RPE65 expressed in COS7 cells at 20 µg

    Secondary
    Alkaline phosphatase-conjugated goat anti-mouse IgG

    Predicted band size : 61 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)
  • ab78036 staining RPE65 in Rhesus monkey retina tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.5% Triton-X and blocked with 4% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody. Counterstained with DAPI. Magnification: 20X.

    See Abreview

  • ab78036 at 1/250 dilution staining RPE65 in frozen tissue sections of retinal pigment epithelium. RPE65 reactivity is restricted to the RPE.
  • ab78036 staining RPE65 in human glioblastoma cell line U87MG by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed with paraformaldehyde, permeabilized using 1% Triton X-100 in PBS, blocked with 0.5% BSA for 20 minutes and then incubated with ab78036 at a 1/50 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse polyclonal, used at a 1/400 dilution.

    Nuclei are stained with DAPI.

    See Abreview

  • Immunofluorescence analysis of ARPE-19 cells, staining RPE65 using ab78036. Cells were treated with PBS, 1 mM EGTA, or 1 mM EGTA plus 10 ng/ml EGF and 20 ng/ml FGF-2 for 1 day.
  • Overlay histogram showing Y79 cells stained with ab78036 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab78036, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Y79 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-RPE65 antibody [401.8B11.3D9] (ab78036)参考文献

This product has been referenced in:
  • Chen HC  et al. Selective activation of p120ctn-Kaiso signaling to unlock contact inhibition of ARPE-19 cells without epithelial-mesenchymal transition. PLoS One 7:e36864 (2012). ICC/IF ; Human . Read more (PubMed: 22590627) »

See 1 Publication for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 22°C
Sample Rhesus monkey Tissue sections (Retina)
Specification Retina
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 May 15 2014

Merci de nous avoir contactés.

Je suis désolé que le remplacement n'ait pas non plus fonctionné.
J'ai demandé à notre service comptabilité de mettre en place un avoir référenc...

Read More

Merci d'avoir pris le temps de remplir notre questionnaire.

Je suis désolé d'apprendre que l'anticorps de remplacement ab78036 n'a pas fonctionné et que vous recontrez, comme avec le ab13826, des problèmes pour la d...

Read More

I'm happy to help! Please let me know if you have any further questions.

Thank you for your enquiry.

The antibody should be OK. We have done internal testing which showed antibodies were stable for at least 7 days at 37 degrees C. I suggest using the antibody as you normally would, ideally using a positive control...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (glioblastoma cell line from rat)
Specification glioblastoma cell line from rat
Fixative Paraformaldehyde
Permeabilization Yes - 1% Triton X 100 in PBS
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Feb 07 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human glioblastoma cell line U87MG)
Specification human glioblastoma cell line U87MG
Fixative Paraformaldehyde
Permeabilization Yes - 1% Triton X 100 in PBS
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 0.5%
Username

Abcam user community

Verified customer

提交于 Feb 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human glioblastoma cell line)
Specification human glioblastoma cell line
Fixative Paraformaldehyde
Permeabilization Yes - 0,1% Triton X 100 in PBS
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 23 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (glioblastoma cell line C6)
Specification glioblastoma cell line C6
Fixative Paraformaldehyde
Permeabilization Yes - 0,1% Triton X 100 in PBS
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jan 23 2012

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