5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236
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More Files
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
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
Flow Cytometry analysis of 5-hmC expression in HEK293 cells using 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat# MAB15116). The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with Cat# MAB15116 (anti-5-hmC, Blue) or with a negative control antibody (RM105, Red).
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.
Enzyme-linked Immunoabsorbent Assay
Direct ELISA of mouse brain genomic DNA using 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat# MAB15116). The plate was directly coated with different concentrations of genomic DNA isolated from mouse brain tissue. 1 ug/mL or 3 ug/mL of Cat# MAB15116 was used as the primary antibody, and a HRP 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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Specification
Product Description
Rabbit recombinant monoclonal antibody raised against 5-hydroxymethylcytosine (5-hmC).
Antibody Species
Rabbit
Immunogen
Original antibody is raised against 5-hydroxymethylcytosine conjugated with BSA.
Sequence
N/A
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.
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Applications
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
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
Flow Cytometry analysis of 5-hmC expression in HEK293 cells using 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat# MAB15116). The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with Cat# MAB15116 (anti-5-hmC, Blue) or with a negative control antibody (RM105, Red).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.Enzyme-linked Immunoabsorbent Assay
Direct ELISA of mouse brain genomic DNA using 5-hydroxymethylcytosine (5-hmC) monoclonal antibody, clone RM236 (Cat# MAB15116). The plate was directly coated with different concentrations of genomic DNA isolated from mouse brain tissue. 1 ug/mL or 3 ug/mL of Cat# MAB15116 was used as the primary antibody, and a HRP 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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