![Western Blot: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Subcellular localization of BEST1 and surface Ca2+-dependent Cl- current in patient-derived iPSC-RPEs. Western blots show similar BEST1 expression levels in WT and patient-derived iPSC-RPEs. Each sample was from one cell lysis (BEST1 and beta-actin, RPE65 and CRALBP were on two gels, respectively). Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/29914), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Western-Blot-NB100-355-img0013.jpg "Western Blot: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Subcellular localization of BEST1 and surface Ca2+-dependent Cl- current in patient-derived iPSC-RPEs. Western blots show similar BEST1 expression levels in WT and patient-derived iPSC-RPEs. Each sample was from one cell lysis (BEST1 and beta-actin, RPE65 and CRALBP were on two gels, respectively). Image collected and cropped by CiteAb from the following publication (//elifesciences.org/articles/29914), licensed under a CC-BY license.")
| Reactivity | Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, XpSpecies Glossary |
| Applications | WB, Simple Western, Flow, Func, ICC/IF, IHC, IP, CyTOF-ready, ICC/IF, KO |
| Clone | 401.8B11.3D9 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Concentration | 1.0 mg/ml |
| Immunogen | This RPE65 Antibody (401.8B11.3D9) was developed against bovine RPE65 microsomal membrane proteins. [UniProt# Q28175] |
| Localization | Cytoplasmic, Microsomeal and Cell Membrane |
| Marker | Retinal Pigment Epithelium Marker |
| Isotype | IgG1 Kappa |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | RPE65 |
| Purity | Protein G purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| Application Notes | For Western blot, this antibody has been validated in lysates of bovine RPE membrane and COS7 cells transfected with human RPE65, and in both samples, the antibody recognized a band at ~65 kDa, representing RPE65 protein. Note:- Hamel et al. have reported that in cultured bovine RPE cells, the levels of RPE65 are undetectable in WB after day 14, however, the mRNA levels are detectable by Northern for at least 7 weeks (PMID: 8340400). The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. This antibody is CyTOF ready. Simple Western reported by an internal validation. Separated by Size |
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| Theoretical MW | 65 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Storage | Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. |
| Buffer | PBS |
| Preservative | 0.02% Sodium Azide |
| Concentration | 1.0 mg/ml |
| Purity | Protein G purified |
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| Images | Ratings | Applications | Species | Date | Details | ||||||||||
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reviewed by:
Verified Customer |
IP | Bovine and Human | 05/06/2019 |
Summary
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reviewed by:
Fabio Marongiu |
ICC | Human | 08/29/2017 |
Summary
Comments
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reviewed by:
Verified Customer |
IF | Other | 03/23/2016 |
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reviewed by:
Verified Customer |
IHC-Fr | Other | 12/12/2014 |
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reviewed by:
Girish Kumar Srivastava |
IF | Other | 10/06/2014 |
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reviewed by:
Girish Kumar Srivastava |
IF | Human | 04/30/2014 |
Summary
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reviewed by:
Verified Customer |
WB | Human | 12/06/2010 |
Summary
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Secondary Antibodies |
Isotype Controls |
Research Areas for RPE65 Antibody (NB100-355)Find related products by research area.
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Using RPE65 as a tool to investigate ocular gene therapies While not life threatening, blindness and retinal disease are profoundly debilitating and greatly affect quality of life. Understandably, gene therapy has been subject to controversy given it’s potential effects on the rest of our cellular ... Read full blog post. |
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RPE65: Vision, Blindness and Hope Retinal pigment epithelium-specific 65 kDa protein (RPE65) is an essential vision protein, and so mutations in the RPE65 gene cause blindness. However, clinical trials using gene therapy to treat patients with a defective RPE65 gene suggest that some ... Read full blog post. |
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
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| Verified Customer 05/06/2019 |
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| Application: | IP | |
| Species: | Bovine and Human |
| Fabio Marongiu 08/29/2017 |
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| Application: | ICC | |
| Species: | Human |
| Verified Customer 03/23/2016 |
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| Application: | IF | |
| Species: | Other |
| Gene Symbol | RPE65 |
![Immunocytochemistry/Immunofluorescence: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Expression of eye-specific markers in the induced eye-like structures induced from lignin-added ES cells. (a) Higher-magnification image of the RPE like structure induced from ESCs after 12-day culture. (b) Immunostaining of eye- like structures. Eye- like structures induced from ESCs after 12-day culture were stained with antibodies against RPE65 (red) and nuclei were stained with DAPI solution (blue). Scale bar: a = 50 um, b,d = 200 um, c,e = 100 um. PLoS One. 2013 Jun 21;8(6):e66376. doi: 10.1371/journal.pone.0066376.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-355-img0014.jpg "Immunocytochemistry/Immunofluorescence: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Expression of eye-specific markers in the induced eye-like structures induced from lignin-added ES cells. (a) Higher-magnification image of the RPE like structure induced from ESCs after 12-day culture. (b) Immunostaining of eye- like structures. Eye- like structures induced from ESCs after 12-day culture were stained with antibodies against RPE65 (red) and nuclei were stained with DAPI solution (blue). Scale bar: a = 50 um, b,d = 200 um, c,e = 100 um. PLoS One. 2013 Jun 21;8(6):e66376. doi: 10.1371/journal.pone.0066376.")
![Immunohistochemistry-Paraffin: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Analysis of FFPE human glioblastoma tissue section using 1:500 dilution of RPE65 [401.8B11.3D9] antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems).](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Immunohistochemistry-Paraffin-NB100-355-img0012.jpg "Immunohistochemistry-Paraffin: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Analysis of FFPE human glioblastoma tissue section using 1:500 dilution of RPE65 [401.8B11.3D9] antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems).")
![Western Blot: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - WB analysis of RPE65 in 20ug lysate of COS7 cells expressing recombinant Human RPE65 (Lane 1) and 5ug of Bovine retinal pigment epithelium membrane fraction (Lane 2). Blot processed for detection with alkaline phosphatase conjugated goat-anti Mouse IgG secondary antibody and NBT/BCIP substrate.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Western-Blot-NB100-355-img0008.jpg "Western Blot: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - WB analysis of RPE65 in 20ug lysate of COS7 cells expressing recombinant Human RPE65 (Lane 1) and 5ug of Bovine retinal pigment epithelium membrane fraction (Lane 2). Blot processed for detection with alkaline phosphatase conjugated goat-anti Mouse IgG secondary antibody and NBT/BCIP substrate.")
![Immunocytochemistry/Immunofluorescence: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - ICC-IF analysis of cultured ARPE19 cells, a spontaneously arising human retinal pigment epithelia cell line - 10 minutes fixation in 4% PFA, 10 minutes permeabilization in PBS containing 0.2% Triton X-100 (PBS-T), 1 hour blocking in 10% normal goat serum containing 1% BSA in PBS-T, 1:100 primary antibody dilution in PBS, ON 4C incubation.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-355-img0006.jpg "Immunocytochemistry/Immunofluorescence: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - ICC-IF analysis of cultured ARPE19 cells, a spontaneously arising human retinal pigment epithelia cell line - 10 minutes fixation in 4% PFA, 10 minutes permeabilization in PBS containing 0.2% Triton X-100 (PBS-T), 1 hour blocking in 10% normal goat serum containing 1% BSA in PBS-T, 1:100 primary antibody dilution in PBS, ON 4C incubation.")
![Immunohistochemistry-Frozen: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Staining of RPE65 in a cryosection of mouse retina tissue using RPE65 antibody (clone 401.8B11.3D9).](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Immunohistochemistry-Frozen-NB100-355-img0007.jpg "Immunohistochemistry-Frozen: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Staining of RPE65 in a cryosection of mouse retina tissue using RPE65 antibody (clone 401.8B11.3D9).")
![Immunohistochemistry: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Zebrafish brain ventricular area labeled with mouse anti-RPE65 (1:100). This image was submitted through a verified customer review.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Immunohistochemistry-NB100-355-img0011.jpg "Immunohistochemistry: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Zebrafish brain ventricular area labeled with mouse anti-RPE65 (1:100). This image was submitted through a verified customer review.")
![Simple Western: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - iPSC-RPE from all donors express RPE marker proteins. iPSC-RPE from the patient, the patients mom, the patients dad, and the three unrelated, unaffected controls all expressed RPE65. The negative control (iPSCs) had no detectable RPE65 protein expression. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29540715/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Simple-Western-NB100-355-img0016.jpg "Simple Western: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - iPSC-RPE from all donors express RPE marker proteins. iPSC-RPE from the patient, the patients mom, the patients dad, and the three unrelated, unaffected controls all expressed RPE65. The negative control (iPSCs) had no detectable RPE65 protein expression. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29540715/) licensed under a CC-BY license.")
![Knockout Validated: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Immunoblotting showing the expression of RPE-specific proteins BEST1 (NB300-164), RPE65 (NB100-355), CRALBP, and the loading control beta-Actin in iPSC-RPE cells. Two gels/blots in the same panel were prepared from the same cell lysate of each PSC-RPE to detect BEST1 + beta-Actin, and RPE65 + CRALBP, respectively. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34061021/) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/RPE65-Antibody-401-8B11-3D9-BSA-Free-Knockout-Validated-NB100-355-img0015.jpg "Knockout Validated: RPE65 Antibody (401.8B11.3D9) - BSA Free [NB100-355] - Immunoblotting showing the expression of RPE-specific proteins BEST1 (NB300-164), RPE65 (NB100-355), CRALBP, and the loading control beta-Actin in iPSC-RPE cells. Two gels/blots in the same panel were prepared from the same cell lysate of each PSC-RPE to detect BEST1 + beta-Actin, and RPE65 + CRALBP, respectively. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34061021/) licensed under a CC-BY license.")
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![Flow Cytometry: Mouse IgG1 Kappa Isotype Control (P3.6.2.8.1) [NBP1-43319] - Analysis of Alexa Fluor (R) 700 conjugate of NBP1-43319. Human buffy coat stained with mouse IgG1 Kappa isotype control. Flow cytometry image submitted by a verified customer review.](https://images.novusbio.com/images/Mouse-IgG1-Kappa-Light-Chain-Isotype-Control-P3-6-2-8-1-Flow-Cytometry-NBP1-43319-img0002.jpg "Flow Cytometry: Mouse IgG1 Kappa Isotype Control (P3.6.2.8.1) [NBP1-43319] - Analysis of Alexa Fluor (R) 700 conjugate of NBP1-43319. Human buffy coat stained with mouse IgG1 Kappa isotype control. Flow cytometry image submitted by a verified customer review.")
