RDH12 anticorps (AA 1-316)
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- Antigène Voir toutes RDH12 Anticorps
- RDH12 (Retinol Dehydrogenase 12 (All-Trans/9-Cis/11-Cis) (RDH12))
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Épitope
- AA 1-316
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Reactivité
- Humain
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Hôte
- Lapin
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Clonalité
- Polyclonal
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Conjugué
- Cet anticorp RDH12 est non-conjugé
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Application
- ELISA, Immunohistochemistry (IHC)
- Réactivité croisée
- Humain
- Purification
- >95%, Protein G purified
- Immunogène
- Recombinant Human Retinol dehydrogenase 12 protein (1-316AA)
- Isotype
- IgG
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- Indications d'application
- Recommended dilution: IHC:1:20-1:200,
- Restrictions
- For Research Use only
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- by
- Palczewski Lab, Center For Translational Vision Research, UC Irvine
- No.
- #104484
- Date
- 23.03.2023
- Antigène
- RDH12
- Numéro du lot
- E0629A
- Application validée
- Immunohistochemistry
- Contrôle positif
Retina cryosection from B6 Albino (B6(Cg)-Tyrc-2J/J) animal
- Contrôle négative
Retina cryosection from B6 Albino (B6(Cg)-Tyrc-2J/J) animal
No primary antibody
- Conclusion
Passed. Presence of specific signal in the RPE cell layer was considered as indication of specific immunoreactivity using the RDH12 antibody ABIN7167836.
- Anticorps primaire
- ABIN7167836
- Anticorps secondaire
- donkey anti-rabbit AF647-conjugated antibody (Abcam, 150075)
- Full Protocol
- Collect eyes from mice and fix with paraformaldehyde 4% (Electron Microscopy Sciences, 15710) in 1x PBS for 30 min at RT.
- Cryoprotection with sucrose series:
- Wash in 10% sucrose in 1x PBS.
- Immerse in 10% sucrose in 1x PBS for 30 min at RT.
- Wash in 20% sucrose in 1x PBS.
- Immerse in 20% sucrose in 1x PBS for 30 min RT.
- Wash in 30% sucrose in 1x PBS.
- 30% sucrose ON at 4°C.
- Embed eyes in OCT compound (Tissue-Tek O.C.T. Compound, 4583).
- Cut retinal sections at a thickness of 12 μm on a cryostat.
- Air dry sections for 15 min at RT, store at -80°C until use.
- Bring sections to RT and rehydrate in 1x PBS for 1 h.
- Incubate sections in blocking buffer (1x PBS, 3% BSA (Sigma-Aldrich, A7030), 3% Donkey serum (Sigma-Aldrich, S30-100ML), 0.1% Triton X-100 (Sigma-Aldrich, X100-500ML)) for 1 h at RT.
- Incubate sections with primary rabbit anti-RDH12 antibody (antibodies-online, ABIN7167836, lot E0629A) diluted 1:50 in blocking buffer ON at RT. Include a no primary antibody negative controls. Additionally, counterstaing with primary mouse anti-CRALBP antibody (Thermo Fisher Scientific, MA1-813).
- Incubate sections with secondary AF647-conjugated donkey anti-rabbit antibody (Abcam, Ab150075) or AF488-conjugated donkey anti-mouse antibody (Thermo Fisher Scientific, A32766) diluted 1:500 in blocking buffer for 1 h at RT.
- Rinse sections once with 1x PBS, 0.1% Triton X-100 for 5 min at RT.
- Incubate sections in 1x DAPI (Thermo Fisher Scientific, 62248) in 1x PBS, 0.1% Triton X-100 for 15 min at RT.
- Rinse sections 3x with 1x PBS, 0.1% Triton X-100 for 5 min at RT.
- Mount sections in VECTASHIELD® HardSet™ Antifade Mounting Medium (Vector Laboratories, H-1400) mounting medium.
- Acquire images with a fluorescence microscope and appropriate filter settings.For the validation purposes Keyence BZ-X800E fluorescence microscope was used with following filters: BZ-X DAPI for DAPI, BZ-X GFP for AF488, BZ-X Cy5 for AF647. Images were taken at 10x and 40x magnification.
- Notes
Experiment involved validation of the specificity of 4 antibodies against different Rdh proteins: Rdh5 (ABIN7254060), Rdh10 (ABIN7118460), Rdh11 (ABIN966957), and Rdh12 (ABIN7167836). All 4 proteins are important for eye function and highly expressed in neural retina and/or RPE. Validation is based on comparison of each staining with known pattern of expression in the mouse retina. For review highlighting each Rdh localization see PMID20801113.
To aid orientation in the cell layers anti-Cralbp counterstain was included in the staining (Thermo MA1-813). Cralbp (Rlbp1) is highly expressed in RPE and Müller glia cells in mouse retina.
Validation #104484 (Immunohistochemistry)Validation ImagesRetinal sections from the wild-type (B6 albino) mice immunostained with anti-RDH12 antibody ABIN7167836. DAPI staining shows localization of the inner (INL) and outer (ONL) nuclear layer of the mouse retina. Cralbp (Rlbp1) co-staining was used to visualize RPE and Müller glia cells in the retina. Presence of specific signal in the RPE cell layer confirms specific immunoreactivity.
Protocole -
- Format
- Liquid
- Buffer
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Preservative: 0.03 % Proclin 300
Constituents: 50 % Glycerol, 0.01M PBS, PH 7.4 - Agent conservateur
- ProClin
- Précaution d'utilisation
- This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Stock
- -20 °C,-80 °C
- Stockage commentaire
- Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
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Rapid RGR-dependent visual pigment recycling is mediated by the RPE and specialized Müller glia." dans: Cell reports, Vol. 42, Issue 8, pp. 112982, (2023) (PubMed).
: "
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Rapid RGR-dependent visual pigment recycling is mediated by the RPE and specialized Müller glia." dans: Cell reports, Vol. 42, Issue 8, pp. 112982, (2023) (PubMed).
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- Antigène
- RDH12 (Retinol Dehydrogenase 12 (All-Trans/9-Cis/11-Cis) (RDH12))
- Autre désignation
- RDH12 (RDH12 Produits)
- Synonymes
- anticorps wu:fj43a10, anticorps zgc:92430, anticorps A930033N07Rik, anticorps LCA13, anticorps LCA3, anticorps RP53, anticorps SDR7C2, anticorps DSSDR2, anticorps retinol dehydrogenase 12 (all-trans/9-cis/11-cis), anticorps retinol dehydrogenase 12, anticorps Retinol dehydrogenase 12, anticorps RDH12, anticorps rdh12, anticorps MAV_1968, anticorps CC1G_02720, anticorps Bm1_36660, anticorps PTRG_03574, anticorps PTRG_05326, anticorps PTRG_08067, anticorps LOC100282710, anticorps LOC100285880, anticorps LOC100226769, anticorps Rdh12
- Sujet
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Background: Exhibits an oxidoreductive catalytic activity towards retinoids. Most efficient as an NADPH-dependent retinal reductase. Displays high activity toward 9-cis and all-trans-retinol. Also involved in the metabolism of short-chain aldehydes. No steroid dehydrogenase activity detected. Might be the key enzyme in the formation of 11-cis-retinal from 11-cis-retinol during regeneration of the cone visual pigments.
Aliases: All trans and 9 cis retinol dehydrogenase antibody, All-trans and 9-cis retinol dehydrogenase antibody, LCA 3 antibody, LCA13 antibody, LCA3 antibody, RDH 12 antibody, RDH12 antibody, RDH12_HUMAN antibody, Retinol dehydrogenase 12 (all trans/9 cis/11 cis) antibody, Retinol dehydrogenase 12 all trans and 9 cis antibody, Retinol dehydrogenase 12 antibody, RP53 antibody, SDR7C2 antibody, Short chain dehydrogenase/reductase family 7C member 2 antibody
- UniProt
- Q96NR8
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