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5-Methylcytosine 抗体

The 兔 多克隆 anti-5-Methylcytosine antibody is suitable to detect 5-Methylcytosine. It has been validated for WB, FACS, IF (p) 和 IHC (p). The antibody has been independently validated.
产品编号 ABIN1385081
发货至: 中国
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Quick Overview for 5-Methylcytosine 抗体 (ABIN1385081)

抗原

5-Methylcytosine

适用

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宿主

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克隆类型

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多克隆

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非结合性

应用范围

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Western Blotting (WB), Flow Cytometry (FACS), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
  • 交叉反应

    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
  • 应用备注

    IHC-P 1:100-500
    IF(IHC-P) 1:50-200

    限制

    仅限研究用
  • 生效 #104610 (Immunohistochemistry)
    '独立验证'标志
    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

    '独立验证'标志
    Validation Images
    Full Methods
    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
  • 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
  • 抗原

    5-Methylcytosine

    别名

    5 MethylCytosine

    物质类

    Chemical

    背景

    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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