Dot Blot (DB)
(Dot blot of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).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 antibody at 0.2 ug/ml.)
Mouse anti-Human 5-Hydroxymethylcytosine Antibody
5-Hydroxymethylcytosine (5-hmC) antibody (rAb)(Clone RM236)
Reactivity
Human
Applications
Dot Blot, Immunofluorescence, Immunohistochemistry
Purity
Protein A Chromatography
Synonyms
5-Hydroxymethylcytosine; 5-Hydroxymethylcytosine (5-hmC) antibody (rAb)(Clone RM236); anti-5-Hydroxymethylcytosine antibody
Host
Mouse
Reactivity
Human
Isotype
IgG
Clone Number
RM236
Purity/Purification
Protein A Chromatography
Form/Format
PBS with 50% glycerol, 1% BSA, and 0.09% sodium azide. Sodium azide is highly toxic.
Concentration
1ug/ul (varies by lot)
Applicable Applications for anti-5-Hydroxymethylcytosine antibody
Dot Blot (DB), Immunofluorescence (IF), Immunohistochemistry (IHC)
Application Notes
DB: 0.2-1ug/ml
IHC: 0.1-1ug/ml
ICC: 0.5-2ug/ml
MeDIP: 0.2-2ug/ml
IHC: 0.1-1ug/ml
ICC: 0.5-2ug/ml
MeDIP: 0.2-2ug/ml
Antibody Type
Recombinant Antibody
Immunogen
BSA-conjugated 5-hydroxymethylcytosine.
Preparation and Storage
Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20 degree C for up to 2 years. Keep all reagents on ice when not in storage.1ug/ul
Dot Blot (DB)
(Dot blot of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).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 antibody at 0.2 ug/ml.)
Dot Blot (DB)
(Dot blot of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).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 antibody at 0.2 ug/ml.)
Testing Data
(MeDIP of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).MeDIP was performed using 5-Hydroxymethylcytosine antibody (Clone 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 1 ug 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.)
Testing Data
(MeDIP of 5-Hydroxymethylcytosine (5-hmC) antibody (rAb).MeDIP was performed using 5-Hydroxymethylcytosine antibody (Clone 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 1 ug 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.)
Testing Data
(Immunofluoresence staining of HeLa cells using 5-Hydroxymethylcytosine antibody (Clone RM236) at 0.5ug/mL (red). Actin filaments are labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (?20 degree C) before treatment with 2 N HCl for 30 min at 37 degree C to denature DNA.)
Testing Data
(Immunofluoresence staining of HeLa cells using 5-Hydroxymethylcytosine antibody (Clone RM236) at 0.5ug/mL (red). Actin filaments are labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (?20 degree C) before treatment with 2 N HCl for 30 min at 37 degree C to denature DNA.)
Related Product Information for anti-5-Hydroxymethylcytosine antibody
Short Description: Our 5-Hydroxymethylcytosine (5-hmC) antibody (rAb)(Clone RM236) was raised in a Rabbit host. It has been validated for use in Dot blot, Immunofluorescence and Immunohistochemistry; it has been shown to react with Human samples, but the sequence is not species specific so it should react with a wide range of sample types.
Background: DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. 5-methylcytosine is a modified base that is found in the DNA of plants and vertebrates. A second type of DNA methylation exists, 5-hydroxymethylcytosine (5-hydroxy methylcytosine, 5-hmC). This results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of cytosine oxygenases. This antibody was developed specifically to distinguish 5-hydroxymethylcytosine from 5-methylcytosine as conventional methods (enrichment by antibody or methyl DNA binding protein, enzymatic digestion and bisulfite sequencing) cannot do so. It is possible that 5-hydroxymethylcytosine (5-hmC) represents a pathway to demethylate DNA, as 5-hydroxymethylcytosine is repaired as mismatched DNA and replaced with unmethylated cytosine.
Background: DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. 5-methylcytosine is a modified base that is found in the DNA of plants and vertebrates. A second type of DNA methylation exists, 5-hydroxymethylcytosine (5-hydroxy methylcytosine, 5-hmC). This results from the enzymatic conversion of 5-methylcytosine into 5-hydroxymethylcytosine by the TET family of cytosine oxygenases. This antibody was developed specifically to distinguish 5-hydroxymethylcytosine from 5-methylcytosine as conventional methods (enrichment by antibody or methyl DNA binding protein, enzymatic digestion and bisulfite sequencing) cannot do so. It is possible that 5-hydroxymethylcytosine (5-hmC) represents a pathway to demethylate DNA, as 5-hydroxymethylcytosine is repaired as mismatched DNA and replaced with unmethylated cytosine.
Product Categories/Family for anti-5-Hydroxymethylcytosine antibody
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Product Notes
The 5-Hydroxymethylcytosine (Catalog #AAA389509) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The 5-Hydroxymethylcytosine (5-hmC) antibody (rAb)(Clone RM236) reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's 5-Hydroxymethylcytosine can be used in a range of immunoassay formats including, but not limited to, Dot Blot (DB), Immunofluorescence (IF), Immunohistochemistry (IHC). DB: 0.2-1ug/ml IHC: 0.1-1ug/ml ICC: 0.5-2ug/ml MeDIP: 0.2-2ug/ml. Researchers should empirically determine the suitability of the 5-Hydroxymethylcytosine for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "5-Hydroxymethylcytosine, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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