JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB214728

Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236]

5

(1 Review)

|

(43 Publications)

Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (ab214728) is a rabbit monoclonal antibody detecting 5-hydroxymethylcytosine (5-hmC) in Flow Cytometry, IP, IHC-P, ICC/IF, ELISA, Dot Blot.

- Recombinant format for unrivalled batch-batch consistency
- Over 20 publications

View Alternative Names

5-hmC

7 Images
Immunocytochemistry/ Immunofluorescence - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

Immunocytochemical staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells using 0.5 μg/mL ab214728 (red). Actin filaments was labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (−20 °C) before treatment with 2 N HCl for 30 minutes at 37°C to denature the DNA.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

Immunohistochemical analysis of paraffin-embedded human brain tissue labeling 5-hydroxymethylcytosine (5-hmC) with ab214728 at 1 μg/mL.

Flow Cytometry - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • Flow Cyt

Supplier Data

Flow Cytometry - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

Flow Cytometry analysis of 5-hmC expression in HEK293 cells using ab214728. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with ab214728 (Blue) or with a negative control antibody (Red).

MeDIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • MeDIP

Supplier Data

MeDIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

hMeDIP was performed using anti-5-hmC antibody (RM236) at a 10 : 1 DNA : Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input.

ELISA - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • ELISA

Supplier Data

ELISA - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

Direct ELISA of mouse brain genomic DNA using ab214728. The plate was directly coated with different concentrations of genomic DNA isolated from mouse brain tissue. 1 ug/mL or 3 ug/mL of ab214728 was used as the primary antibody and an HRP conjugated anti-rabbit IgG as the secondary antibody.

ELISA - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • ELISA

Supplier Data

ELISA - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

ELISA of single stranded DNA using ab214728 in a serial dilution. The plate was coated with streptavidin and then biotinylated single stranded unmethylated DNA, 5-Methylcytosine (5-mC) DNA, and 5-Hydroxymethylcytosine (5-hmC) DNA. Secondary antibody : alkaline phosphatase conjugated anti-rabbit IgG. ``

Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)
  • Dot

Supplier Data

Dot Blot - Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (AB214728)

Dot blot of double stranded DNA using ab214728 at 0.2 μg/mL. The membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with ab214728.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

RM236

Isotype

IgG

Carrier free

No

Applications

Flow Cyt, IHC-P, MeDIP, Dot, ICC/IF, ELISA

applications

Specificity

ab214728 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.

style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "ELISA" : {"fullname" : "ELISA", "shortname":"ELISA"}, "MeDIP" : {"fullname" : "MeDIP", "shortname":"MeDIP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Modified Nucleic Acid": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "0.2-1 g/mL", "Dot-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "0.1-1 µg/mL", "IHCP-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "0.5-2 µg/mL", "ICCIF-species-notes": "<p></p>", "ELISA-species-checked": "testedAndGuaranteed", "ELISA-species-dilution-info": "0.1-1 µg/mL", "ELISA-species-notes": "<p></p>", "MeDIP-species-checked": "testedAndGuaranteed", "MeDIP-species-dilution-info": "0.2-2 µg/mL", "MeDIP-species-notes": "<p></p>", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "1-5 µg/mL", "FlowCyt-species-notes": "<p></p>" } } }
style
left-column

Product details

What is this antibody validated in?
Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (ab214728) is a rabbit recombinant monoclonal antibody and is validated for use in Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), ELISA, Dot Blot. Trusted by the scientific community
Anti-5-hydroxymethylcytosine (5-hmC) [RM236] (ab214728) was first used in a scientific publication in 2016 and has been cited over 20 times in peer-reviewed journals.

Other related products
We have a range of other formats of antibody clone [RM236] also available for your convenience: ab214728, ab319976, Carrier free - ab323582
style
left-column

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
Preservative: 0.09% Sodium azide Constituents: PBS, 50% Glycerol (glycerin, glycerine), 1% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle
style
left-column

Supplementary information

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

5-hydroxymethylcytosine (5-hmC) also known as hydroxymethylcytosine represents a significant epigenetic modification in DNA. This target with a mass of approximately 63.3 emerges through DNA demethylation processes where ten-eleven translocation (TET) enzymes oxidize 5-methylcytosine (5-mC). It is found largely in the brain where it shows high levels of expression but it can also appear in other tissues to a lesser extent. The modification is important in regulating gene expression and maintaining cellular identity.
Biological function summary

5-hydroxymethylcytosine plays a critical role in epigenetic regulation and serves as an intermediary in the active DNA demethylation process. It is not part of a stable complex but interacts transiently during epigenetic reconfiguration. The presence of 5-hmC marks regulatory regions of genes and is involved in transcriptional activation or repression depending on the cellular context. Its levels are dynamic influenced by various developmental stages and environmental conditions.

Pathways

This DNA modification is an important player in pathways related to cellular differentiation and development. It is integrally involved in the DNA demethylation pathway where TET enzymes mediate its formation from 5-methylcytosine. This pathway also involves other modifications leading to cytosine turnover linking 5-hmC to broader chromatin remodeling processes. Proteins interacting with 5-hmC include TET enzymes and other DNA maintenance proteins which highlight its role in maintaining genomic stability.

Aberrant 5-hmC levels show connection to several pathological states including cancer and neurological disorders. Altered hydroxymethylation patterns often associate with tumorigenesis particularly in gliomas where the misregulation of 5-hmC accompanies cancer progression. Neurological disorders like Rett syndrome also exhibit altered levels of 5-hmC implicating its role in neurodevelopmental processes. In these diseases mutations in proteins such as TET enzymes link to disrupted 5-hmC levels indicating its critical involvement in both oncogenesis and neurodevelopmental regulation.
style
left-column
style
left-column

Product protocols

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

Target data

See full target information 5-hydroxymethylcytosine (5-hmC)
style
left-column

Publications (43)

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

Frontiers in molecular neuroscience 18:1628860 PubMed40747242

2025

Tet3-mediated DNA demethylation is essential for maintaining the dedifferentiation capacity of mammalian Müller glia.

Applications

Unspecified application

Species

Unspecified reactive species

Erick J Martinez-Colin,Ivonne Lezama,Lenin D Ochoa-de la Paz,Monica Lamas

Cell death & disease 16:553 PubMed40701963

2025

H3K14 lactylation exacerbates neuronal ferroptosis by inhibiting calcium efflux following intracerebral hemorrhagic stroke.

Applications

Unspecified application

Species

Unspecified reactive species

Tingting Sun,Jian-Nan Zhang,Ting Lan,Lei Shi,Liye Hu,Liyan Yan,Chao Wei,Lisha Hei,Weihua Wu,Zhaoli Luo,Meng Liu,Xingmei Ren,Yamei Wang,Yabin Lu,Peipei Wang,Chenguang Zhang,Liheng Bian,Xingquan Zhao,Fei Yang,Qian Li

Scientific reports 15:15633 PubMed40325038

2025

Immunohistochemical detection of 5-hydroxymethylcytosine as a prognostic biomarker in non-WNT/Non-SHH medulloblastomas.

Applications

Unspecified application

Species

Unspecified reactive species

Han-Guang Zhao,Si-Kang Ren,Wen-Tao Zhou,Zhi-Ming Liu,Lei-Ming Wang,Yong-Ji Tian,Fu Zhao

Heliyon 10:e40957 PubMed39759294

2025

Transcriptomic effects of alginate hydrogel applied to the production of bovine embryos.

Applications

Unspecified application

Species

Unspecified reactive species

Giuliana de A Ferronato,Paola M da S Rosa,Alessandra Bridi,Angélica Camargo Dos Santos,Ricardo P Nociti,Marcos Roberto Chiaratti,Felipe Perecin,Flávio V Meirelles,Juliano R Sangalli,Juliano C da Silveira

Aging cell 24:e14458 PubMed39722450

2024

DNMT3a Deficiency Contributes to Anesthesia/Surgery-Induced Synaptic Dysfunction and Cognitive Impairment in Aged Mice.

Applications

Unspecified application

Species

Unspecified reactive species

Peilin Cong,Xinwei Huang,Qian Zhang,Mengfan He,Hanxi Wan,Qianqian Wu,Huanghui Wu,Yuxin Zhang,Chun Cheng,Li Tian,Lize Xiong

iScience 27:111382 PubMed39687026

2024

Mission SpaceX CRS-16 RRRM-1 space flight induced skin genomic plasticity via an epigenetic trigger.

Applications

Unspecified application

Species

Unspecified reactive species

Kanhaiya Singh,Priyanka Verma,Rajneesh Srivastava,Yashika Rustagi,Manishekhar Kumar,Sumit S Verma,Sujit Mohanty,Afshin Beheshti,Liz Warren,Chandan K Sen

eLife 13: PubMed39642055

2024

Maternal obesity may disrupt offspring metabolism by inducing oocyte genome hyper-methylation via increased DNMTs.

Applications

Unspecified application

Species

Unspecified reactive species

Shuo Chao,Jun Lu,Li-Jun Li,Hong-Yan Guo,Kuipeng Xu,Ning Wang,Shu-Xian Zhao,Xiao-Wen Jin,Shao-Ge Wang,Shen Yin,Wei Shen,Ming-Hui Zhao,Gui-An Huang,Qing-Yuan Sun,Zhao-Jia Ge

BMC pharmacology & toxicology 25:42 PubMed39010179

2024

DNA methylation, but not microRNA expression, is affected by in vitro THC exposure in bovine granulosa cells.

Applications

Unspecified application

Species

Unspecified reactive species

Sabrina Floccari,Reem Sabry,Laurie Choux,Michael S Neal,Jibran Y Khokhar,Laura A Favetta

Journal of experimental botany 75:3849-3861 PubMed38652039

2024

Non-canonical bases differentially represented in the sex chromosomes of the dioecious plant Silene latifolia.

Applications

Unspecified application

Species

Unspecified reactive species

Marcel Hubinský,Roman Hobza,Marta Starczak,Daniel Gackowski,Zdeněk Kubát,Tomáš Janíček,Lucie Horáková,Jose Luis Rodriguez Lorenzo

Epigenetics 19:2337142 PubMed38583183

2024

Vitamin C enhances co-localization of novel TET1 nuclear bodies with both Cajal and PML bodies in colorectal cancer cells.

Applications

Unspecified application

Species

Unspecified reactive species

Nour El Osmani,Corinne Prévostel,Laurence Picque Lasorsa,Mohammad El Harakeh,Zeina Radwan,Hiba Mawlawi,Marwan El Sabban,Margret Shirinian,Zeina Dassouki
View all publications
style
left-column
style
left-column
style
right-column

Alternative Products

Primary Antibodies

AB106918

Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade

ChIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (AB106918)

  • 3
  • 2

Complementary Products

Primary Antibodies

AB106918

Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade

ChIP - Anti-5-hydroxymethylcytosine (5-hmC) antibody [AB3/63.3] - ChIP Grade (AB106918)

  • 3
  • 2
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB9485

Anti-GAPDH antibody - Loading Control

BOND RX™ Validated|Lab Essentials

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (AB9485)

  • 5
  • 88
Primary Antibodies

AB10805

Anti-5-methylcytosine (5-mC) antibody [33D3]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (AB10805)

  • 5
  • 12

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