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Key Documents

MABN2438

Sigma-Aldrich

Anti-RPE65 Antibody, clone KPSA1

Synonym(s):

All-trans-retinyl-palmitate hydrolase, EC:3.1.1.64, Lutein isomerase, Meso-zeaxanthin isomerase, Retinal pigment epithelium-specific 65 kDa protein, Retinoid isomerohydrolase, Retinol isomerase

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

KPSA1, monoclonal

species reactivity

bovine, human, mouse

packaging

antibody small pack of 100

technique(s)

direct ELISA: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
western blot: suitable

isotype

IgG1κ

epitope sequence

C-terminal

Protein ID accession no.

UniProt accession no.

Gene Information

human ... RPE65(6121)

Specificity

Clone KPSA1 is a mouse monoclonal antibody that detects RPE65. It targets an epitope within 23 amino acid from the C-terminal region.

Immunogen

Recombinant full-length Human RPE65 expressed in Sf9 cells.

Application

Quality Control Testing

Evaluated by Western Blotting in Bovine retina microsomal preparation.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected RPE65 in Bovine retina microsomal preparation.

Tested Applications

Western Blotting Analysis: A representative lot detected RPE65 in Western Blotting applications (Golczak, M., et al. (2010). J Biol Chem. 285(13):9667-9682; Banskota, S., et al. (2022). Cell. 185(2):250-265.e16).

Immunoaffinity Purification: A representative lot was used for purification of crossed-linked RPE65.(Golczak, M., et al. (2010). J Biol Chem. 285(13):9667-9682).

Immunohistochemistry Applications: A representative lot detected RPE65 in Immunohistochemistry applications (Amengual, J., et al. (2014). Hum Mol Genet. 23(20):5402-17).

ELISA Analysis: A representative lot detected RPE65 in ELISA applications (Golczak, M., et al. (2010). J Biol Chem. 285(13):9667-9682).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Retinoid isomerohydrolase (UniProt: Q16518; also known as EC:3.1.1.64, All-trans-retinyl-palmitate hydrolase, Lutein isomerase, Meso-zeaxanthin isomerase, Retinal pigment epithelium-specific 65 kDa protein, Retinol isomerase) is encoded by the RPE65 gene (Gene ID: 6121) in human. Retinol isomerase is a lipid-anchor protein that is attached to the membrane when palmitoylated and is present in soluble form when unpalmitoylated. It is shown to be critical for retinoid cycle and is involved in regeneration of 11-cis-retinal, the chromophore of rod and cone opsins. It catalyzes the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol that is further oxidized by 11-cis retinol dehydrogenase to 11-cis-retinal for use as visual chromophore. It can also catalyze the isomerization of lutein to meso-zeaxanthin an eye-specific carotenoid. The soluble form is reported to binds vitamin A (all-trans-retinol), making it available for lecithin retinol acyltransferase (LRAT) processing to all-trans-retinyl ester. The membrane form, palmitoylated by LRAT, binds all-trans-retinyl esters, making them available for isomerohydrolase processing to all-cis-retinol. The soluble form can be regenerated by transferring its palmitoyl groups onto 11-cis-retinol, a reaction catalyzed by LRAT. It undergoes light-dependent intracellular transport to become more concentrated in the central region of the retina pigment epithelium cells. Mutations in RPE65 gene are known to cause Retinitis pigmentosa that is characterized by retinal pigment deposits visible on fundus examination and primary loss of rod photoreceptor cells followed by secondary loss of cone photoreceptors. (Ref.: Golczak, M., et al. (2010). J. Biol. Chem. 285(13); 9667-9682; Moiseyev, G., et al. (2005). Proc. Natl. Acad. Sci. USA. 102(35):12413-12418).

Physical form

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl and 0.05% sodium azide.

Reconstitution

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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