Rabbit monoclonal antibody raised against 5-hydroxymethylcytosine (5-hmC).
Immunogen:
5-hydroxymethylcytosine conjugated with BSA.
Host:
Rabbit
Specificity:
This antibody reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA.
Form:
Liquid
Purification:
Protein A affinity purification
Isotype:
IgG
Recommend Usage:
Dot Blot (0.2-1 ug/mL)ELISA (0.1-1 ug/mL)Immunocytochemistry (0.5-2 ug/mL)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) (0.1-1 ug/mL)Hydroxymethylated DNA immunoprecipitation (0.2-2 ug/mL)The optimal working dilution should be determined by the end user.
Storage Buffer:
In PBS (50% glycerol, 1% BSA, 0.09% sodium azide)
Storage Instruction:
Store at -20°C.Aliquot to avoid repeated freezing and thawing.
Note:
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Immunohistochemical staining (Formalin-fixed paraffin-embedded sections) of human brain with 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat # MAB15116).
Immunocytochemistry
Immunocytochemistry staining of HeLa cells with 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat # MAB15116) (Red). Actin filaments was labeled with fluorescein phalloidin (Green).
Enzyme-linked Immunoabsorbent Assay
ELISA analysis of single stranded DNA with 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat # MAB15116). The plate was coated with streptavidin and then biotinylated single stranded unmethylated DNA, 5-Methylcytosine (5-mC) DNA, and 5-Hydroxymethylcytosine (5-hmC) DNA. A serial dilution of MAB15116 was used as the primary antibody, and an alkaline phosphatase conjugated anti-rabbit IgG as the secondary antibody.
Dot Blot
Dot blot analysis of double stranded DNA with 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat # MAB15116). 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 MAB15116.
Methylated DNA Immunoprecipitation
Hydroxymethylated DNA immunoprecipitation (hMeDIP) analysis of 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat # MAB15116) 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.
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