Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPRNCI-184-128] to N4-acetylcytidine (ac4C) - BSA and Azide free
  • Suitable for: Dot blot, Northern Blot, RIP

Overview

  • Product name

    Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free
    See all N4-acetylcytidine (ac4C) primary antibodies

  • Description

    Rabbit monoclonal [EPRNCI-184-128] to N4-acetylcytidine (ac4C) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Dot blot, Northern Blot, RIPmore details

  • Immunogen

    Chemical/ Small Molecule. This information is proprietary to Abcam and/or its suppliers.

  • 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

    Ab253039 is the carrier-free version of ab252215. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab253039 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPRNCI-184-128

  • Isotype

    IgG

Images

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

    This data was developed using the same antibody clone in a different buffer formulation (ab252215).

    Dot blot analysis of N4-acetylcytidine (ac4C) labeled with ab252215 at 1/200 dilution.

    Lane 1: HeLa total RNA
    Lane 2: ac4C RNA oligo
    Lane 3: C RNA oligo

    Goat Anti-Rabbit IgG H&L (HRP) Peroxidase conjugated (ab97051) at 1/20,000 dilution was used as secondary antibody.

     

    Ac4C and C oligos (without biotin labelled) used in the figure are from collaborator. We also tested biotin-C oligo and it showed negative (Lane3).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

    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 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039) Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 3C.; doi: 10.1021/acschembio.7b00734.

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039) Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 4C.; 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Dot Blot - Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039) Image from Sinclair WR et al., ACS Chemical Biology., 12 (12), 2922-2926. Fig 4F.; 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252215).

  • Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)
    Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] - BSA and Azide free (ab253039)

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