RDH10 antibody

Catalog No. ABIN7118460

Cited in 1 publication. The Rabbit Polyclonal anti-RDH10 antibody (ABIN7118460) specifically detects RDH10 in WB, IHC, ELISA and IP. The antibody is reactive with Human, Rat and Mouse samples. Independently validated for use in Immunohistochemistry.

Quick Overview for RDH10 antibody (ABIN7118460)

Target: RDH10 (Retinol Dehydrogenase 10 (All-Trans) (RDH10))

Reactivity: Human, Rat, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This RDH10 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunoprecipitation (IP)

Product Details anti-RDH10 Antibody

Purpose: RDH10 antibody

Purification: Immunogen affinity purified

Purity: ≥95 % as determined by SDS-PAGE

Immunogen: retinol dehydrogenase 10(all-trans)

Isotype: IgG

Application Details

Application Notes: WB: 1:500-1:2000, IP: 1:200-1:1000, IHC: 1:20-1:200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104469 (Immunohistochemistry)

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

No.: #104469

Date: 03/23/2023

Antigen: RDH10

Lot Number: 20221223E

Method validated: Immunohistochemistry

Positive Control:

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

Negative Control:

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

No primary antibody

Notes:

Passed. Presence of specific signal in the RPE cell layer was considered as indication of specific immunoreactivity using the anti-RDH10 antibody ABIN7118460.

Validation Images

Retinal sections from the wild-type (B6 albino) mice immunostained with anti-Rdh10 antibody ABIN7118460. 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.

Full Methods

Primary Antibody: ABIN7118460

Secondary Antibody: 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-RDH10 antibody (antibodies-online, ABIN7118460, lot 20221223E) 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).
• Rinse sections 3 times with 1x PBS, 0.1% Triton X100. Keep negative controls in a separate container.
• 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.

Experimental 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.

Handling

Format: Liquid

Buffer: PBS with 0.02 % sodium azide and 50 % glycerol pH 7.3,

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freeze / thaw cycles.

Storage: -20 °C

Storage Comment: -20°C for 12 months

Expiry Date: 12 months

References for anti-RDH10 antibody (ABIN7118460)

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).

Target Details for RDH10

Target: RDH10 (Retinol Dehydrogenase 10 (All-Trans) (RDH10))

Alternative Name: RDH10

Background:

Synonyms: Retinol dehydrogenase 10|Short chain dehydrogenase/reductase family 16C member 4|RDH10|SDR16C4

Background: Retinol dehydrogenase with a clear preference for NADP. Converts all-trans-retinol to all-trans-retinal. Has no detectable activity towards 11-cis-retinol, 9-cis-retinol and 13-cis-retinol.

Molecular Weight: 39 kDa

Gene ID: 157506

UniProt: Q8IZV5