• Western blot analysis of extracts of various cell lines, using RPE65 antibody (STJ111883) 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% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 5s.
  • Perform microwave antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with IF staining protocol. Immunofluorescence analysis of paraffin-embedded rat retina using RPE65 Rabbit polyclonal antibody (STJ111883) at dilution of 1:300 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunofluorescence analysis of Rat eye cells using RPE65 antibody (STJ111883) at dilution of 1:100. Blue: DAPI for nuclear staining.
  • Perform microwave antigen retrieval with 10 mM citrate buffer pH 6. 0 before commencing with IF staining protocol. Immunofluorescence analysis of paraffin-embedded mouse retina using RPE65 Rabbit polyclonal antibody (STJ111883) at dilution of 1:300 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunofluorescence analysis of mouse eye cells using RPE65 antibody (STJ111883) at dilution of 1:100. Blue: DAPI for nuclear staining.
  • Western blot analysis of lysates from Mouse eye, using RPE65 Rabbit polyclonal antibody (STJ111883) at 1:3000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 30s.

Anti-RPE65 antibody (274-533) (STJ111883)

SKU:
STJ111883

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Host: Rabbit
Applications: WB/IF
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-RPE65 (274-533) is suitable for use in Western Blot and Immunofluorescence research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.02% Sodium Azide, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:1000-1:5000
IF/ICC 1:50-1:200
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: RPE65
Gene ID: 6121
Uniprot ID: RPE65_HUMAN
Immunogen Region: 274-533
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 274-533 of human RPE65 (NP_000320.1).
Immunogen Sequence: NYMDCFESNETMGVWLHIAD KKRKKYLNNKYRTSPFNLFH HINTYEDNGFLIVDLCCWKG FEFVYNYLYLANLRENWEEV KKNARKAPQPEVRRYVLPLN IDKADTGKNLVTLPNTTATA ILCSDETIWLEPEVLFSGPR QAFEFPQINYQKYCGKPYTY AYGLGLNHFVPDRLCKLNVK TKETWVWQEPDSYPSEPIFV SHPDALEEDDGVVLSVVVSP GAGQKPAYLLILNAKDLSE
Tissue Specificity Retina (at protein level). Retinal pigment epithelium specific.
Post Translational Modifications 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).
Function Critical isomerohydrolase in the retinoid cycle involved in regeneration of 11-cis-retinal, the chromophore of rod and cone opsins. Catalyzes the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol which is further oxidized by 11-cis retinol dehydrogenase to 11-cis-retinal for use as visual chromophore. Essential for the production of 11-cis retinal for both rod and cone photoreceptors. Also capable of catalyzing the isomerization of lutein to meso-zeaxanthin an eye-specific carotenoid. 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.
Protein Name Retinoid Isomerohydrolase
All-Trans-Retinyl-Palmitate Hydrolase
Lutein Isomerase
Meso-Zeaxanthin Isomerase
Retinal Pigment Epithelium-Specific 65 Kda Protein
Retinol Isomerase
Database Links Reactome: R-HSA-2453902
Cellular Localisation Cytoplasm
Cell Membrane
Lipid-Anchor
Microsome Membrane
Attached To The Membrane By A Lipid Anchor When Palmitoylated (Membrane Form)
Soluble When Unpalmitoylated
Undergoes Light-Dependent Intracellular Transport To Become More Concentrated In The Central Region Of The Retina Pigment Epithelium Cells
Alternative Antibody Names Anti-Retinoid Isomerohydrolase antibody
Anti-All-Trans-Retinyl-Palmitate Hydrolase antibody
Anti-Lutein Isomerase antibody
Anti-Meso-Zeaxanthin Isomerase antibody
Anti-Retinal Pigment Epithelium-Specific 65 Kda Protein antibody
Anti-Retinol Isomerase antibody
Anti-RPE65 antibody

Information sourced from Uniprot.org

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