Anti-5-Hydroxymethylcytosine antibody, Rabbit monoclonal
SIGMA/SAB5600041 - recombinant, expressed in HEK 293 cells, clone RM236, purified immunoglobulin
Synonym: 5-hmC
Product Type: Chemical
Immunohistochemistry Immunohistochemical staining of formalin fixed and paraffin embedded Human brain tissue sections, using Rabbit monoclonal Anti-5-hmC (clone RM236) antibody.
Immunocytochemistry Immunocytochemical staining of HeLa cells using 0.5mg/mL Anti-5-hmC antibody (RM236) (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 min at 37 °C to denature the DNA.
Dot Blot Dot blot of double stranded DNA using Anti-5-hmC antibody (RM236) antibody. 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.
Immunoprecipitation 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 ELISA of single stranded DNA using Anti-5-hmC antibody (RM236). 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 RM236 was used as the primary antibody, and an alkaline phosphatase conjugated Anti-Rabbit IgG as the secondary antibody.
ELISA Direct ELISA of Mouse brain genomic DNA using Anti-5-hmC antibody (RM236). The plate was directly coated with different concentrations of genomic DNA isolated from Mouse brain tissue. 1 μg/mL or 3 μg/mL of RM236 was used as the primary antibody, and a HRP conjugated Anti-Rabbit IgG as the secondary antibody.
| antibody form | purified immunoglobulin |
| antibody product type | primary antibodies |
| clone | recombinant monoclonal |
| RM236, monoclonal | |
| concentration | ~1 mg/mL |
| form | buffered aqueous glycerol solution |
| isotype | IgG |
| Quality Level | 100  |
| recombinant | expressed in HEK 293 cells |
| shipped in | wet ice |
| species reactivity | human |
| storage temp. | −20°C |
| technique(s) | dot blot: 0.2-1 μg/mL |
| ELISA: 0.1- 1 μg/mL | |
| immunocytochemistry: 0.5-2 μg/mL | |
| immunohistochemistry: 0.1 ug/mL – 1 ug/mL |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
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| Immunogen: | BSA-conjugated 5-hydroxymethylcytosine |
| Physical form: | Solution in phosphate buffered saline containing 50% glycerol, 1% BSA and 0.09% sodium azide. |
| 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. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |