AB251529

Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free

RabMAb
Recombinant
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(1 Publication)

Rabbit Recombinant Monoclonal N4-methylcytidine (m4C) antibody. Carrier free. Suitable for IP, ELISA and reacts with Modified Nucleic Acid samples. Cited in 1 publication.

View Alternative Names

N4-methylcytidine, m4C

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ELISA

Supplier Data

ELISA - Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free (AB251529)

This data was developed using ab211494, the same antibody clone in a different buffer formulation.

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.

Supplier Data

Immunoprecipitation - Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free (AB251529)

This data was developed using ab211494, the same antibody clone in a different buffer formulation.

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 : 5% NFDM/TBST

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

All lanes:

Immunoprecipitation - Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] (<a href='/en-us/products/primary-antibodies/n4-methylcytidine-m4c-antibody-epr-19850-108-ab211494'>ab211494</a>)

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Unconjugated

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR -19850-108

Isotype

IgG

Carrier free

Yes

Applications

IP, ELISA

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Has been developed to discriminate between the modified base N4-methylcytidine (m4C) and the unmodified counterpart Cytidine (C).

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ELISA" : {"fullname" : "ELISA", "shortname":"ELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Modified Nucleic Acid": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "ELISA-species-checked": "testedAndGuaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "<p></p>" } } }

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Product details

ab251529 is the carrier-free version of ab211494.

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

N4-methylcytidine (m4C) is a modified nucleoside found in RNA specifically involved in post-transcriptional modifications. It is sometimes referred to as methylcytidine and its mass is approximately 257.23 Da. You can find m4C expressed in various prokaryotic and eukaryotic systems suggesting its broad role in molecular biology. Notably the methylation process leading to m4C is facilitated by specific methyltransferase enzymes. Because of this enzymatic activity the nucleoside exhibits diverse mechanical functions that influence RNA stability and translation efficiency.

Biological function summary

M4C plays a role in the regulation of gene expression. It is normally observed as part of the ribosomal RNA in both bacterial and mammalian cells indicating its conserved function across species. The methylation at the cytidine position in RNA helps stabilize the RNA structure which contributes to the control of protein synthesis. m4C does not directly form part of any known complex but the modifications it undergoes indirectly affect ribosomal complexes and their function.

Pathways

The modifications introduced by m4C are fundamental to the mRNA translation pathway and RNA processing pathway. The methylated form interacts with ribosomal proteins influencing translational fidelity and efficiency. m4C also associates with RNA polymerase proteins due to its participation in message stability. These interactions position m4C as a pivotal contributor in maintaining cellular homeostasis by regulating protein synthesis through these essential pathways.

M4C modifications relate to certain types of cancers and neurological disorders. Abnormal m4C levels can disrupt normal gene expression patterns contributing to oncogenesis. Furthermore its association with ribosomal processing implicates it in neurodegenerative diseases where protein misfolding is a characteristic feature. Proteins like ribosomal protein S5 and transcription factor E2F1 have connections to m4C's altered functions in these conditions revealing the complex role m4C plays in disease etiology. Understanding m4C's involvement might help in developing targeted therapies for conditions linked to translation deregulation.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information N4-methylcytidine (m4C)

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Oncology letters 18:545-552 PubMed31289526

2019

Lymphoepithelioma-like gastric carcinoma treated with partial gastrectomy: Two case reports.

Applications

Unspecified application

Species

Unspecified reactive species

Huihua Cao,Jun Xie,Yongxiang Qian,Yugang Wu,Zhaoqing Tang

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free

ELISA - Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free (AB251529)

Immunoprecipitation - Anti-N4-methylcytidine (m4C) antibody [EPR -19850-108] - BSA and Azide free (AB251529)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Applications

Immunogen

Specificity

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

Product promise