Recombinant
RabMAb

Recombinant Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] (ab211494)

Overview

  • Product name
    Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108]
    See all N4-methylcytidine (m4C) primary antibodies
  • Description
    Rabbit monoclonal [EPR -19850-108] to N4-methylcytidine (m4C)
  • Host species
    Rabbit
  • Specificity
    Has been developed to discriminate between the modified base N4-methylcytidine (m4C) and the unmodified counterpart Cytidine (C).
  • Tested applications
    Suitable for: IP, ELISAmore details
  • Species reactivity
    Reacts with: Synthetic fragment
  • Immunogen

    Chemical/ Small Molecule corresponding to N4-methylcytidine (m4C).

  • Positive control
    • IP: 5' Biotin-mN.mN.mN.mN.mN.[m4C]*.mN.mN.mN.mN.mN 3' * - phosphorothioate protection. ELISA: Biotinylated m4C-modified oligonucleotides.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR -19850-108
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab211494 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent concentration.

0.2 µg of ab211494 was coated onto 50 µL Dynabeads® sheep-anti-rabbit IgG.

ELISA Use a concentration of 0.005 - 4 µg/ml.

Target

  • Alternative names
    • m4C antibody
    • N4-methylcytidine antibody

Images

  • The IP was performed in a U-bottom non-adsorbing propylene 96-well plate.

    ab211494 (0.2 μg) was coated into Dynabeads® sheep-anti-rabbit IgG (50 μl) for 1h at RT.

    Unmodified/modified oligonucleotides (5 μM) were added to samples containing the antibody/bead complexes and incubated with agitation for 1 hour at RT.

    After washing, Peroxidase-conjugated Streptavidin was incubated at 1/1000 dilution with agitation for 1 hour at RT.

    ECL substrate was then added and the results read in a non-transparent 96-well plate with a digital detector and analyzed using ImageJ.

    Lane 1: Buffer only.

    Lane 2: Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m4C]*.mN.mN.mN.mN.mN 3’

    Lane 3: Unmodified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[C]*.mN.mN.mN.mN.mN 3’

    N - equimolar mixture of (A/U/G/C)

    m - 2’O methyl protection

    * - phosphorothioate protection

     

    Blocking buffer and concentration: 5% NFDM/TBST

    Dilution buffer and concentration: TBST/0.1% Triton X-100/1 mM EDTA

  • Biotinylated m4C (modified) and C (unmodified) oligonucleotides with the below sequence were coated onto wells of a 96 well plate. 

    Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m4C]*.mN.mN.mN.mN.mN 3’

    Unmodified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[C]*.mN.mN.mN.mN.mN 3’

    N - equimolar mixture of (A/U/G/C)
    m - 2’O methyl protection
    * - phosphorothioate protection

    ELISA was performed on 1.0 µg/ml of antigen using ab211494 at a concentration range of 0.005-4.000 µg/ml, followed by Goat Anti-Rabbit IgG, (H+L), alkaline phosphatase conjugated secondary antibody at 1/2500 dilution.

References

ab211494 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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