5-Methylcytosine 抗体
(1 validation)-
- 抗原
- 5-Methylcytosine
- 适用
- 请咨询
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- 非结合性
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应用范围
- Western Blotting (WB), Flow Cytometry (FACS), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
- 交叉反应
- 人
- 交叉反应 （详细）
- 5 MethylCytosine
- 纯化方法
- Purified by Protein A.
- 免疫原
- KLH conjugated 5 MethylCytosine
- 亚型
- IgG
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anti-5-Methylcytosine antibody
适用: 人 MeDIP, IF, DB 宿主: 小鼠 Monoclonal A1 unconjugated
anti-5-Methylcytosine antibody (Biotin)IF, ICC 宿主: 小鼠 Monoclonal 5MC-CD Biotin
anti-5-Methylcytosine antibodyIF, ICC 宿主: 小鼠 Monoclonal 5MC-CD unconjugated
anti-5-Methylcytosine antibody (FITC)IF, ICC 宿主: 小鼠 Monoclonal 5MC-CD FITC
anti-5-Methylcytosine antibodyWB, MeDIP, FACS, IF, DB, ICC, IHC, ELISA 宿主: 小鼠 Monoclonal 33D3 unconjugated
anti-5-Methylcytosine antibodyMeDIP, DB, MeDIP-Seq 宿主: 小鼠 Monoclonal A1 unconjugated
anti-5-Methylcytosine antibodyMeDIP, ICC 宿主: 兔 Polyclonal unconjugated
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- 应用备注
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IHC-P 1:100-500
IF(IHC-P) 1:50-200 - 限制
- 仅限研究用
-
- by
- Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
- No.
- #104610
- 日期
- 2024.07.29
- 抗原
- 5-Methylcytosine
- Lot Number
- 9K04V1
- Method validated
- Immunohistochemistry
- Positive Control
Adult (> 2 months) CD1 mouse brain fixed in 4% paraformaldehyde.
- Negative Control
One control slice for each experimental procedure processed omitting the primary antibody; overnight incubation in diluent solution only.
- Notes
Passed. The antibody works for IHC-P at 1:100 and 1:200 concentrations without microwave antigen retrieval
- Primary Antibody
- ABIN1385081
- Secondary Antibody
- Full Protocol
- Perfuse mice with paraformaldehyde 4% in 0.1 M phosphate buffer pH 7.4 and post-fix of samples in the same fixative for an additional 2 h at RT.
- Wash, dehydrate, and embed samples in paraffin wax.
- Wash several times with 0.01 M PBS.
- Cut the brain with a microtome into 6 µm sections and mount them on glass slides.
- After paraffin removal, sections were incubated for 1 hour at room temperature in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot # 00583) to block non-specific binding sites.
- Incubate sections with the primary antibody anti-5-methyl-cytosine (ABIN1385081; Lot 9K04V1) at 1:50, 1:100, and 1:200 in PBS-BSA-(Sigma, A7906)- PLL (Sigma, P1524) at RT in a humid chamber.
- Wash sections 3x 5 min with 0.01 M PBS.
- Incubate sections with anti-rabbit secondary antibody Alexa Fluor 488, 1:500, in 0.1M PBS (Thermo Fisher Scientific, A11034, lot # 2380031) for 1 h at RT.
- Wash sections 3x 5 min with 0.01 M PBS.
- Specimens were mounted in Fluoroshield (Sigma, F6182, lot # MKCB0153V).
- Images were acquired with a Leica DM 6000B fluorescence microscope with a digital camera at 40x magnification.
- Experimental Notes
生效 #104610 (Immunohistochemistry)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![5-methyl-cytosine immunoreactive neuronal nuclei in the mouse forebrain. A: a group of immunoreactive nuclei in the deep layers of the cerebral cortex (original magnification 20x); B: at higher magnification (40x), the positive 5-methyl-cytosine sites are visible in the form of a punctate reaction. Primary antibody dilution: 1:100. No microwave.]()
5-methyl-cytosine immunoreactive neuronal nuclei in the mouse forebrain. A: a group of immunoreactive nuclei in the deep layers of the cerebral cortex (original magnification 20x); B: at higher magnification (40x), the positive 5-methyl-cytosine sites are visible in the form of a punctate reaction. Primary antibody dilution: 1:100. No microwave.
Full Methods -
- 状态
- Liquid
- 浓度
- 1 μg/μL
- 缓冲液
- 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
- 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
- 4 °C,-20 °C
- 储存方法
- Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
- 有效期
- 12 months
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- 抗原
- 5-Methylcytosine
- 别名
- 5 MethylCytosine
- 物质类
- Chemical
- 背景
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Synonyms: 5-Methyl Cytidine, 5 m C, 5 mC, 5 me C, 5 Me Cytidine, 5 MeCyd, 5 Methyl Cytidine, 5 MethylCytosine, 5-mC, 5-Me Cytidine, 5-Methyl-Cytosine, 5-MethylCytidine.
Background: Cytidine is a nucleoside formed by a cytosine attached to a ribose ring via a beta-N1-glycosidic bond. DNA is methylated on cytidines by DNA methylases (DNMTs)to generate 5-methylcytidine (5-mC), a potent epigenetics marker and regulator of gene expression. The reverse reaction (cytidine demethylation) starts with its oxidation to hydroxymethyl- (5-hmC), formyl- (5-fC), and carboxy- (5-caC) cytidine. Several enzymes, including the Tet family of proteins have been implicated in cytidine demethylation.
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