RDH12 Antikörper (AA 1-316)

Produktnummer ABIN7167836

Produktdetails anti-RDH12 Antikörper

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

Bindungsspezifität: AA 1-316

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser RDH12 Antikörper ist unkonjugiert

Applikation: ELISA, Immunohistochemistry (IHC)

Kreuzreaktivität: Human

Aufreinigung: >95%, Protein G purified

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

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: Recommended dilution: IHC:1:20-1:200,

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104484 (Immunohistochemistry)

by: Palczewski Lab, Center For Translational Vision Research, UC Irvine

No.: #104484

Datum: 23.03.2023

Antigen: RDH12

Chargennummer: E0629A

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

Retina cryosection from B6 Albino (B6(Cg)-Tyrc-2J/J) animal

Negativkontrolle:

Retina cryosection from B6 Albino (B6(Cg)-Tyrc-2J/J) animal

No primary antibody

Bewertung:

Passed. Presence of specific signal in the RPE cell layer was considered as indication of specific immunoreactivity using the RDH12 antibody ABIN7167836.

Validierungsbilder

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.

Protokoll

Primärantikörper: ABIN7167836

Sekundärantikörper: 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.

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Buffer: Preservative: 0.03 % Proclin 300
Constituents: 50 % Glycerol, 0.01M PBS, PH 7.4

Konservierungsmittel: ProClin

Vorsichtsmaßnahmen: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: -20 °C,-80 °C

Informationen zur Lagerung: Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Referenzen für anti-RDH12 Antikörper (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." in: Cell reports, Vol. 42, Issue 8, pp. 112982, (2023) (PubMed).

Details zu RDH12

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

Andere Bezeichnung: RDH12 (RDH12 Produkte)

Synonyme: wu:fj43a10 antikoerper, zgc:92430 antikoerper, A930033N07Rik antikoerper, LCA13 antikoerper, LCA3 antikoerper, RP53 antikoerper, SDR7C2 antikoerper, DSSDR2 antikoerper, retinol dehydrogenase 12 (all-trans/9-cis/11-cis) antikoerper, retinol dehydrogenase 12 antikoerper, Retinol dehydrogenase 12 antikoerper, RDH12 antikoerper, rdh12 antikoerper, MAV_1968 antikoerper, CC1G_02720 antikoerper, Bm1_36660 antikoerper, PTRG_03574 antikoerper, PTRG_05326 antikoerper, PTRG_08067 antikoerper, LOC100282710 antikoerper, LOC100285880 antikoerper, LOC100226769 antikoerper, Rdh12 antikoerper

Hintergrund:

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