RDH11 抗体
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Quick Overview for RDH11 抗体 (ABIN966957)
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See all RDH11 抗体适用
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应用范围
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anti-Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11) (AA 22-318) antibody
RDH11 适用: 人 WB, ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11) (AA 24-318) antibodyRDH11 适用: 人 WB, ELISA 宿主: 小鼠 Monoclonal 1H6 unconjugated
anti-Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11) (C-Term) antibodyRDH11 适用: 人, 大鼠, 豚鼠, 马, Pig, 兔, Bat, 猴 WB 宿主: 兔 Polyclonal unconjugated
anti-Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11) antibodyRDH11 适用: 人, 大鼠 WB 宿主: 兔 Polyclonal unconjugated
anti-Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11) (C-Term) antibodyRDH11 适用: 人, 小鼠, 大鼠, 豚鼠, 马, Pig, 兔, Cow, 犬 WB 宿主: 兔 Polyclonal unconjugated
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限制
- 仅限研究用
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- by
- Palczewski Lab, Center For Translational Vision Research, UC Irvine
- No.
- #104470
- 日期
- 2023.03.23
- 抗原
- RDH11
- Lot Number
- 0618
- 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-RDH11 antibody ABIN966957.
- Primary Antibody
- ABIN966957
- 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-RDH11 antibody (antibodies-online, ABIN966957, lot 0618) 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.
- 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.
生效 #104470 (Immunohistochemistry)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Retinal sections from the wild-type (B6 albino) mice immunostained with anti-RDH11 antibody ABIN966957. 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.]()
Retinal sections from the wild-type (B6 albino) mice immunostained with anti-RDH11 antibody ABIN966957. 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 -
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储存条件
- 4 °C
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- RDH11 (Retinol Dehydrogenase 11 (All-Trans/9-Cis/11-Cis) (RDH11))
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别名
- RDH11
抗原
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