RPE65 Recombinant Rabbit Monoclonal Antibody [JM61-51]
Catalog# ET1703-82
RPE65 Recombinant Rabbit Monoclonal Antibody [JM61-51]
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WB
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IHC-P
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IF-Tissue
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IHC-Fr
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
RPE65 Recombinant Rabbit Monoclonal Antibody [JM61-51]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human RPE65 aa 441-484 / 533.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Tissue, IHC-Fr, IP
Target Molecular Weight
Predicted band size: 61 kDa
Positive Control
Mouse eyeball tissue lysate, Rat eyeball tissue lysate, rat eyeball tissue, mouse eyeball tissue.
Conjugation
unconjugated
Clone Number
JM61-51
RRID
Reactivity Data
| WB | IHC-P | IF-Tissue | IHC-Fr | IP | |
|---|---|---|---|---|---|
| Human | |||||
| Mouse |
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| Rat |
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Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:1,000
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IF-Tissue
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1:500
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IHC-Fr
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1:500
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IP
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1-2μg/sample
Target
Function
The retinal pigment epithelium (RPE) is a monolayer simple epithelium in proximity to the outer surface of the retinal photoreceptor cells. Retinal pigment epithelium-specific protein (RPE65) is a 65kDa protein belonging to the beta-carotene dioxygenase family. This protein is important in 11-cis retinal production as well as in visual pigment regeneration. RPE65 is attached to the membrane by a lipid anchor when palmitoylated (membrane form) and soluble when unpalmitoylated. The soluble form of the protein binds vitamin A. Defects in RPE65 causes autosomal dominant retinitis pigmentosa and/or Leber congenital amaurosis type 2.
Background References
1. Zhou J et al. Correlations between Photodegradation of Bisretinoid Constituents of Retina and Dicarbonyl Adduct Deposition. J Biol Chem 290:27215-27 (2015).
2. Sukseree S et al. Tyrosinase-Cre-Mediated Deletion of the Autophagy Gene Atg7 Leads to Accumulation of the RPE65 Variant M450 in the Retinal Pigment Epithelium of C57BL/6 Mice. PLoS One 11:e0161640 (2016).
Sequence Similarity
Belongs to the carotenoid oxygenase family.
Tissue Specificity
Retina (at protein level). Retinal pigment epithelium specific.
Post-translational Modification
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) (By similarity).
Subcellular Location
Cytoplasm. Cell membrane.
Synonyms
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
ExpandImages
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Application: Immunofluorescence (IHC-Fr)
Species: Mouse
Tissue: Retina
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-82, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Application: Immunofluorescence (IHC-Fr)
Species: Rat
Tissue: Retina
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-82, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature. -
Western blot analysis of RPE65 on different lysates with Rabbit anti-RPE65 antibody (ET1703-82) at 1/2,000 dilution.
Lane 1: Mouse eyeball tissue lysate
Lane 2: Rat eyeball tissue lysate
Lysates/proteins at 40 µg/Lane.
Predicted band size: 61 kDa
Observed band size: 61 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-82) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Retina
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1703-82, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Retina
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1703-82, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature. -
Application: Immunofluorescence (IF-tissue)
Species: Mouse
Tissue: Retina
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: ET1703-82, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"