RPE65 Antibody (5X1F6)

Novus Biologicals | Catalog # NBP3-16840

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 5X1F6 expressed in HEK293
View all RPE65 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide corresponding to a sequence within amino acids 400-500 of human RPE65 (Q16518). TIWLEPEVLFSGPRQAFEFPQINYQKYCGKPYTYAYGLGLNHFVPDRLCKLNVKTKETWVWQEPDSYPSEPIFVSHPDALEEDDGVVLSVVVSPGAGQKPA

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

61 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.

Scientific Data Images for RPE65 Antibody (5X1F6)

Western Blot: RPE65 Antibody (5X1F6) [NBP3-16840]

Western Blot: RPE65 Antibody (5X1F6) [NBP3-16840]

Western Blot: RPE65 Antibody (5X1F6) [NBP3-16840] - Analysis of extracts of Rat eye, using RPE65 Rabbit mAb (NBP3-16840) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 3min.
Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840] - Immunofluorescence analysis of mouse eye using RPE65 Rabbit mAb (NBP3-16840) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840] - Rat retina using RPE65 Rabbit mAb (NBP3-16840) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry-Paraffin: RPE65 Antibody (5X1F6) [NBP3-16840] - Mouse retina using RPE65 Rabbit mAb (NBP3-16840) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840]

Immunohistochemistry: RPE65 Antibody (5X1F6) [NBP3-16840] - Immunofluorescence analysis of rat eye using RPE65 Rabbit mAb (NBP3-16840) at dilution of 1:100 (40x lens). Blue: DAPI for nuclear staining.
RPE65 Antibody (5X1F6)

Immunocytochemistry/ Immunofluorescence: RPE65 Antibody (5X1F6) [NBP3-16840] -

Immunocytochemistry/ Immunofluorescence: RPE65 Antibody (5X1F6) [NBP3-16840] - Immunofluorescence analysis of paraffin-embedded rat eye using RPE65 Rabbit mAb at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.

Applications for RPE65 Antibody (5X1F6)

Application
Recommended Usage

ELISA

Recommended starting concentration is 1 μg/mL

Immunocytochemistry/ Immunofluorescence

1:100 - 1:1000

Immunohistochemistry

1:100 - 1:1000

Immunohistochemistry-Paraffin

1:100 - 1:1000

Western Blot

1:500 - 1:1000

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

PBS (pH 7.3), 50% glycerol, 0.05% BSA

Preservative

0.02% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: RPE65

RPE65 (retinal pigment epithelium-specific 65 kDa protein) was first described in 1991 and is a key isomerase in RPE that is largely localized to the smooth endoplasmic reticulum (1,2). The RPE65 gene is located on chromosome 1p31 and the protein is synthesized as 533 amino acids in length with a theoretical molecular weight of 61 kDa, although appearing near 65 kDa in SDS-PAGE (1,3). RPE65 serves an important enzymatic function in the visual cycle in converting all-trans-retinyl esters into 11-cis-retinal (1-6). In the visual cycle, light activates rhodopsin and other visual pigments in the photoreceptor cells (rods and cones), eventually generating all-trans-retinol which is transported to the RPE and converted to the vitamin A-derived chromophore 11-cis-rentinal via RPE65 and is then diffused back to the photoreceptor to continue the cycle (1-6). Although it is still unclear, some studies have suggested that S-palmitoylation post-translational modification of RPE65 is responsible for the stability and anchoring to the RPE membrane and therefore functioning enzymatic activity (1,2).

Given its essential role in the vision cycle, it is understandable that mutations in RPE65 are associated with a variety of inherited retinal dystrophies (1, 3-6). Leber Congenital Amaurosis (LCA) and retinitis pigmentosa (RP) are two of the most common retinal dystrophies associated with bi-allelic RPE65 gene mutations (5,6). In 2017 the FDA approved an in vivo gene therapy for treatment of RPE65-associated diseases (5,6). The drug Voretigene Neparvovec, also called Luxturna, is delivered sub-retinally and transduces RPE cells with cDNA encoding for normal RPE65 to help restore vision (5,6). There are several promising completed and ongoing clinical trials for treating RPE65-associated diseases using gene replacement therapy (5).

References

1. Kiser, P. D., & Palczewski, K. (2010). Membrane-binding and enzymatic properties of RPE65. Progress in retinal and eye research. https://doi.org/10.1016/j.preteyeres.2010.03.002

2. Uppal, S., Poliakov, E., Gentleman, S., & Redmond, T. M. (2019). RPE65 Palmitoylation: A Tale of Lipid Posttranslational Modification. Advances in experimental medicine and biology. https://doi.org/10.1007/978-3-030-27378-1_88

3. Redmond T. M. (2009). Focus on Molecules: RPE65, the visual cycle retinol isomerase. Experimental eye research. https://doi.org/10.1016/j.exer.2008.07.015

4. Saari J. C. (2016). Vitamin A and Vision. Sub-cellular biochemistry. https://doi.org/10.1007/978-94-024-0945-1_9

5. Miraldi Utz, V., Coussa, R. G., Antaki, F., & Traboulsi, E. I. (2018). Gene therapy for RPE65-related retinal disease. Ophthalmic genetics. https://doi.org/10.1080/13816810.2018.1533027

6. Apte R. S. (2018). Gene Therapy for Retinal Degeneration. Cell. https://doi.org/10.1016/j.cell.2018.03.021

Alternate Names

All-trans-retinyl-palmitate hydrolase, BCO family, member 3, BCO3, EC 3.1.1.64, EC:3.1.1.64, EC:5.3.3.22, LCA2, lutein isomerase, meso-zeaxanthin isomerase, mRPE65, p63, RBP-binding membrane protein, rd12, retinal pigment epithelium specific protein 65, Retinal pigment epithelium-specific 65 kDa protein, retinal pigment epithelium-specific protein 65kDa, retinitis pigmentosa 20 (autosomal recessive), retinoid isomerohydrolase, Retinol isomerase, RP20, RPE65, RPE65, retinoid isomerohydrolase, sRPE65

Gene Symbol

RPE65

Additional RPE65 Products

Product Documents for RPE65 Antibody (5X1F6)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for RPE65 Antibody (5X1F6)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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FAQs

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