Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] ab252215 is a rabbit monoclonal antibody that is used in N4-acetylcytidine (ac4C) dot blot. Suitable for modified nucleic acid samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
Dot | |
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Modified Nucleic Acid | Tested |
Species | Dilution info | Notes |
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Species Modified Nucleic Acid | Dilution info 1/200 - 1/1000 | Notes - |
Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] ab252215 is a rabbit monoclonal antibody that is used in N4-acetylcytidine (ac4C) dot blot. Suitable for modified nucleic acid samples.
- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPRNCI-184-128
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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 monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
N4-acetylcytidine (ac4C) a modified nucleoside occurs in various types of RNA like tRNA rRNA and mRNA. The molecular mass of ac4C roughly totals 255 daltons. It is expressed ubiquitously across many organisms including humans yeast and bacteria. This modification also referred to sometimes as an 'RNA acetylation mark' contributes to the stability and codon-anticodon interactions in tRNA with an important role in maintaining the structural integrity of rRNA components.
N4-acetylcytidine acts to enhance translation fidelity which is essential for accurate gene expression. It ensures the stabilization of RNA structures and impacts the efficiency of translation. This modification often involves an enzyme complex including acetyltransferases which work by transferring the acetyl group from acetyl-CoA to the cytidine residue. Proper functioning and balance of ac4C modifications in RNA depend on these enzymes ensuring that the RNA molecule remains functional and effective in protein synthesis.
N4-acetylcytidine takes part in translation and RNA processing pathways. It works alongside RNA polymerase complexes and other enzymes that modify RNA. The modification of ac4C enhances the initiation and elongation phases of translation. Within these pathways it interacts with proteins such as RNA acetylating enzymes and ribosomal proteins to fine-tune these processes ensuring that RNA synthesis and subsequent protein production are precise and efficient.
N4-acetylcytidine shows links to conditions related to translation errors and cellular stress responses. Aberrant levels of ac4C have connections to cancer wherein dysregulated translation and misfolded protein formation occur. Defective ac4C modification might lead to neurodegeneration where accurate protein synthesis is compromised. Proteins like NAT10 a known RNA acetyltransferase interact with ac4C in these pathological contexts indicating the importance of maintaining balanced ac4C levels for cellular health and disease prevention.
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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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20,000 dilution was used as secondary antibody.
Ac4C oligo (without biotin labelled) used in the figure was provided by a collaborator.
Competitive RNA Dot Blot - Recombinant Anti-N4-acetylcytidine (ac4C) antibody [EPRNCI-184-128] (ab252215)
Primary antibody dilution: 1 ug/ml
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated
Secondary antibody dilution: 1/20,000
Blocking buffer and dilution buffer: AdvanBlock™ Chemi Blocking buffer
Input: HeLa total RNA 500ng per Dot
Competitive nucleosides:
Lane A: ac4C
Lane B: unmodified C
Exposure time: 137 seconds
This blot was developed using a higher sensitivity ECL substrate
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.
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