RDH12 anticorps (AA 1-316)

N° du produit ABIN7167836

Cet validé indépendamment anticorps Lapin Polyclonal détecte spécifiquement RDH12 dans ELISA et IHC. Il présente une réactivité envers Humain et a été mentionné dans 1 publication.

Aperçu rapide pour RDH12 anticorps (AA 1-316) (ABIN7167836)

Antigène: RDH12 (Retinol Dehydrogenase 12 (All-Trans/9-Cis/11-Cis) (RDH12))

Reactivité: Humain

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp RDH12 est non-conjugé

Application: ELISA, Immunohistochemistry (IHC)

Détail du produit anti-RDH12 anticorps

Épitope: AA 1-316

Réactivité croisée: Humain

Purification: >95%, Protein G purified

Immunogène: Recombinant Human Retinol dehydrogenase 12 protein (1-316AA)

Isotype: IgG

Information d'application

Indications d'application: Recommended dilution: IHC:1:20-1:200,

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104484 (Immunohistochemistry)

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.

Validation Images

Retinal 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

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.

Stockage

Format: Liquid

Buffer: 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.

Références pour anticorps anti-RDH12 (ABIN7167836)

Tworak, Kolesnikov, Hong, Choi, Luu, Palczewska, Dong, Lewandowski, Brooks, Campello, Swaroop, Kiser, Kefalov, Palczewski: "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).

Détails sur RDH12

Antigène: RDH12 (Retinol Dehydrogenase 12 (All-Trans/9-Cis/11-Cis) (RDH12))

Autre désignation: RDH12

Sujet:

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