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RDH10 抗体

RDH10 适用: 人, 小鼠, 大鼠 WB, IHC, ELISA, IP 宿主: 兔 Polyclonal unconjugated
产品编号 ABIN7118460
发货至: 中国
  • 抗原 See all RDH10 抗体
    RDH10 (Retinol Dehydrogenase 10 (All-Trans) (RDH10))
    适用
    • 28
    • 5
    • 5
    • 4
    • 4
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    人, 小鼠, 大鼠
    宿主
    • 28
    克隆类型
    • 28
    多克隆
    标记
    • 9
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This RDH10 antibody is un-conjugated
    应用范围
    • 13
    • 13
    • 13
    • 10
    • 9
    • 8
    • 5
    • 3
    • 2
    • 1
    Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunoprecipitation (IP)
    纯化方法
    Immunogen affinity purified
    纯度
    ≥95 % as determined by SDS-PAGE
    免疫原
    retinol dehydrogenase 10(all-trans)
    亚型
    IgG
  • 应用备注
    WB: 1:500-1:2000, IP: 1:200-1:1000, IHC: 1:20-1:200
    限制
    仅限研究用
  • 生效 #104469 (Immunohistochemistry)
    '独立验证'标志
    by
    Palczewski Lab, Center For Translational Vision Research, UC Irvine
    No.
    #104469
    日期
    2023.03.23
    抗原
    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.

    '独立验证'标志
    Validation Images
    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.

  • 状态
    Liquid
    缓冲液
    PBS with 0.02 % sodium azide and 50 % glycerol pH 7.3,
    储存液
    Sodium azide
    注意事项
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    储存条件
    -20 °C
    储存方法
    -20°C for 12 months (Avoid repeated freeze / thaw cycles.)
    有效期
    12 months
  • 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).

  • 抗原
    RDH10 (Retinol Dehydrogenase 10 (All-Trans) (RDH10))
    别名
    RDH10 (RDH10 产品)
    背景
    Synonyms: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.
    分子量
    39 kDa
    基因ID
    157506
    UniProt
    Q8IZV5
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