Recombinant
RabMAb

Recombinant Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] (ab252215)

Overview

  • Product name
    Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128]
    See all N4-acetylcytidine (ac4C) primary antibodies
  • Description
    Rabbit monoclonal [EPRNCI-184-128] to N4-acetylcytidine (ac4C)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Dot blot, Northern Blot, RIPmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to N4-acetylcytidine (ac4C) conjugated to keyhole limpet haemocyanin.

  • Positive control
    • Dot blot: Synthetic E. coli tRNA Met probe containing ac4C; synthetic Beta globin RNA probe containing ac4C; Total RNA purified from wild type HeLa cells.
  • 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
    EPRNCI-184-128
  • Isotype
    IgG

Applications

Our Abpromise guarantee covers the use of ab252215 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
Dot blot 1/1000.
Northern Blot 1/500.

Immuno Northern blot. (1 μg/mL).

(see Figure 1 in Arango D. et al., 2018. Acetylation of Cytidine in mRNA Promotes Translation Efficiency. Cell 175(7):1872-1886).

RIP 1/50.

(10 μg/mL).

(see Figure 3 in Arango D. et al., 2018. Acetylation of Cytidine in mRNA Promotes Translation Efficiency. Cell 175(7):1872-1886).

Images

  • Dot blot analysis of N4-acetylcytidine (ac4C) labeled with either ab252215 at 1/1000 dilution or with methylene blue.

    Lane 1: Total RNA purified from wild-type Hela cells.
    Lane 2: Total RNA purified from NAT10 knockout HeLa cells

    Goat Anti-Rabbit IgG H&L (HRP) at 1/100000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times: Left panel, 2 minutes. Right panel, 30 seconds.

    Knock out of NAT10 (human acetyltransferase) can dramatically reduce ac4C level.

    This image was kindly provided by Dr. Jordan Meier, NCI, USA.

    This antibody was cited in PMID: 29039931 and 30449621.

  • Dot blot analysis of N4-acetylcytidine (ac4C) labeled with ab252215 at 1/1000 dilution.

    Lane 1: Synthetic wild type Beta globin RNA probe.
    Lane 2: Synthetic Beta globin RNA probe containing ac4C.
    Lane 3: Synthetic Beta globin RNA probe containing m5C.
    Lane 4: Beta globin RNA probe containing hm5C.

    Goat Anti-Rabbit IgG H&L (HRP) at 1/100000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 2 seconds.

    This antibody shows no cross reactivity with other RNA modifications.

    This image was kindly provided by Dr. Jordan Meier, NCI, USA.

    This antibody was cited in PMID: 29039931 and 30449621.

  • Dot blot analysis of N4-acetylcytidine (ac4C) labeled with ab252215 at 1/1000 dilution.

    Lane 1: Synthetic wild type E. coli tRNA Met probe.
    Lane 2: Synthetic E. coli tRNA Met probe containing ac4C.
    Lane 3: Synthetic E. coli tRNA Met probe containing ac4C pre-treated with 50mM hydroxylamine.

    Goat Anti-Rabbit IgG H&L (HRP) at 1/100000 dilution was used as the secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 2 seconds.

    Hydroxylamine removes the electrophilic acetamide of ac4C.

    This image was kindly provided by Dr. Jordan Meier, NCI, USA.

    This antibody was cited in PMID: 29039931 and 30449621.

  • Dot blot analysis demonstrates concentration-dependent detection of in vitro transcribed ac4C-containing RNAs. The antibody detects RNA probes synthesized in the presence of ac4CTP, but not CTP.

    Refer to PMID 29039931 

    Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 3C.; doi: 10.1021/acschembio.7b00734.

  • Dot blot analysis showing effect of hydroxylamine on ac4C in RNA using anti-ac4C antibody. Reaction conditions: 50 mM hydroxylamine, Tris at pH 8.0, 65 °C, 1 hour. Hydroxylamine reduced the detection of ac4C.

    Refer to PMID 29039931

     

    Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 4C.; doi: 10.1021/acschembio.7b00734.

  • Dot blot analysis of total RNA by anti-ac4C found a significantly decreased signal in hydroxylamine-treated samples compared to untreated samples.

    Refer to PMID 29039931

     

    Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 4F.; doi: 10.1021/acschembio.7b00734.

References

This product has been referenced in:
See all 2 Publications for this product

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