Recombinant Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236] (ab214728)
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: ELISA, MeDIP, Flow Cyt, Dot blot, IHC-P, ICC/IF
- Reacts with: Species independent
Overview
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Product name
Anti-5-hydroxymethylcytosine (5-hmC) antibody [RM236]
See all 5-hydroxymethylcytosine (5-hmC) primary antibodies -
Description
Rabbit monoclonal [RM236] to 5-hydroxymethylcytosine (5-hmC) -
Host species
Rabbit -
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. -
Tested applications
Suitable for: ELISA, MeDIP, Flow Cyt, Dot blot, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to 5-hydroxymethylcytosine (5-hmC) conjugated to Bovine Serum Albumin (BSA).
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Positive control
- IHC-P: Human brain tissue. ICC/IF: HeLa cells.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 1% BSA, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
RM236 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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ELISA |
Use a concentration of 0.1 - 1 µg/ml.
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MeDIP |
Use a concentration of 0.2 - 2 µg/ml.
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Flow Cyt |
Use a concentration of 1 - 5 µg/ml.
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Dot blot |
Use a concentration of 0.2 - 1 µg/ml.
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IHC-P |
Use a concentration of 0.1 - 1 µg/ml.
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ICC/IF |
Use a concentration of 0.5 - 2 µg/ml.
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Notes |
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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. |
ICC/IF
Use a concentration of 0.5 - 2 µg/ml. |
Target
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Relevance
5-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. -
Alternative names
- 5-hmC antibody
Images
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Immunohistochemical analysis of paraffin-embedded human brain tissue labeling 5-hydroxymethylcytosine (5-hmC) with ab214728 at 1 μg/mL.
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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.
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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.
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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.
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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
.
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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.
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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).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (21)
ab214728 has been referenced in 21 publications.
- Zachou K et al. Altered DNA methylation pattern characterizes the peripheral immune cells of patients with autoimmune hepatitis. Liver Int 42:1355-1368 (2022). PubMed: 35108441
- Shen Y et al. Loss of 5-hydroxymethylcytosine as a Poor Prognostic Factor for Primary Testicular Diffuse Large B-cell Lymphoma. Int J Med Sci 19:225-232 (2022). PubMed: 35165508
- Kong FC et al. A novel epigenetic marker, Ten-eleven translocation family member 2 (TET2), is identified in the intractable epileptic brain and regulates ATP binding cassette subfamily B member 1 (ABCB1) in the blood-brain barrier. Bioengineered 13:6638-6649 (2022). PubMed: 35235761
- Zhang J et al. Metformin Regulates TET2 Expression to Inhibit Endometrial Carcinoma Proliferation: A New Mechanism. Front Oncol 12:856707 (2022). PubMed: 35480097
- Cao LH et al. Food Allergy-Induced Autism-Like Behavior is Associated with Gut Microbiota and Brain mTOR Signaling. J Asthma Allergy 15:645-664 (2022). PubMed: 35603013