Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [RM236] to 5-hydroxymethylcytosine (5-hmC)
- Suitable for: ICC, ELISA, MeDIP, Flow Cyt, Dot blot, IHC-P
Product nameAnti-5-hydroxymethylcytosine (5-hmC) antibody [RM236]
See all 5-hydroxymethylcytosine (5-hmC) primary antibodies
DescriptionRabbit monoclonal [RM236] to 5-hydroxymethylcytosine (5-hmC)
Specificityab214728 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.
Tested applicationsSuitable for: ICC, ELISA, MeDIP, Flow Cyt, Dot blot, IHC-Pmore details
Chemical/ Small Molecule corresponding to 5-hydroxymethylcytosine (5-hmC) conjugated to Bovine Serum Albumin (BSA).
- IHC-P: Human brain tissue. ICC/IF: HeLa cells.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped 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 bufferPreservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 1% BSA, PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab214728 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use a concentration of 0.5 - 2 µg/ml.|
|ELISA||Use a concentration of 0.1 - 1 µg/ml.|
|MeDIP||Use a concentration of 0.2 - 2 µg/ml.|
|Flow Cyt||Use a concentration of 1 - 5 µg/ml.|
|Dot blot||Use a concentration of 0.2 - 1 µg/ml.|
|IHC-P||Use a concentration of 0.1 - 1 µg/ml.|
Relevance5-Hydroxymethylcytosine (5-hmC) is a modified base form of cytosine recently found in Human/mouse brain and inembryonic stem cells. This DNA pyrimidine nitrogen base can be generated by oxidation of 5-methylcytosine, a reaction mediated by the ten-eleven translocation (TET) family of the 5-mC hydroxylases. The function of this base is still not elucidated but it is believed to play an important role in switching genes on and off.
- 5-hmC antibody
Immunohistochemical analysis of paraffin-embedded human brain tissue labeling 5-hydroxymethylcytosine (5-hmC) with ab214728 at 1 μg/mL.
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.
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.
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 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 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.
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).
ab214728 has been referenced in 6 publications.
- Wu T et al. Characterization of global 5-hydroxymethylcytosine in pediatric posterior fossa ependymoma. Clin Epigenetics 12:19 (2020). PubMed: 31992357
- Loktev SS & Ogneva IV DNA Methylation of Mouse Testes, Cardiac and Lung Tissue During Long-Term Microgravity Simulation. Sci Rep 9:7974 (2019). PubMed: 31138883
- Tong M et al. 5-Hydroxymethylcytosine as a potential epigenetic biomarker in papillary thyroid carcinoma. Oncol Lett 18:2304-2309 (2019). PubMed: 31452729
- Zhong J & Xu W Characterization of DNA hydroxymethylation in the hypothalamus of elderly mice with post-operative cognitive dysfunction. Exp Ther Med 18:4002-4010 (2019). PubMed: 31641381
- MacKay H et al. DNA methylation in AgRP neurons regulates voluntary exercise behavior in mice. Nat Commun 10:5364 (2019). PubMed: 31792207
- Almeida LO et al. SET oncoprotein accumulation regulates transcription through DNA demethylation and histone hypoacetylation. Oncotarget 8:26802-26818 (2017). PubMed: 28460463