Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig2_hmC_dotblot.png)
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig1_5hmC_ELISA.png)
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig3_Brain_5hmc.png)
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig5_DirectELISA_hmC_e.png)
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig4_hMeDIP_hmC_e.png)
![Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody; [Clone RM236]; 5′-hmC; 31-1111-00](/upload/image/products/Fig6_ICC_hmC_e.png)
Key features and details
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Product Details
| Product Name | Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody, Clone RM236 |
| Product Description | Rabbit monoclonal to 5-hydroxymethylcytosine (5-hmC); 5'-hmC |
| Catalog No. | 31-1111-00 |
| Clone Name | RM236 |
| Product Data Sheet | Anti-5-hydroxymethylcytosine (5-hmC) Rabbit Monoclonal Antibody Data Sheet RM236 |
| Specificity | RM236 reacts to 5-hydroxymethylcytosine in both single-stranded and double-stranded DNA. No cross reactivity with non-methylated cytosine and methylcytosine in DNA. |
| Immunogen | BSA-conjugated 5-hydroxymethylcytosine. |
| Application | Hydroxymethylated DNA immunoprecipitation (hMeDIP) Immunohistochemistry (IHC) Immunocytochemistry (ICC) ELISA Dot Blot (DB) Flow Cytometry (FC) |
| Species Reactivity | All |
| Conjugate | None |
| Intended Use | Research Use Only |
| . | |
| Form | Liquid |
| Storage Instructions | Store at -20.0°C |
| Stability | Stable for 1 Year at -20.0°C from date of receipt. |
| Storage Buffer | 50% Glycerol/PBS with 1% BSA and 0.09% sodium azide |
| Volume/Size | 50 µg |
| Concentration | 1.0 mg/mL |
| Purity | Protein A affinity purified from an animal origin–free culture supernatant |
| Clone Type | Monoclonal |
| Isotype | Rabbit IgG |
| Usage | Dot Blot: 0.2 µg /mL - 1 µg /mL; ELISA: 0.1 ug/mL - 1 ug/mL; IHC: 0.1 µg/mL - 1 µg/mL; ICC: 0.5 µg/mL - 2 µg/mL; hMeDIP: 0.2 µg/mL - 2 µg/mL; FC:1 µg/mL - 5 µg/mL |
| References for 5-hydroxymethylcytosine (5-hmC) antibody [RM236] | Siref AB et al. Diagnostic utility of dual 5-hmC/Melan-A immunohistochemistry in differentiating nodal nevus from metastatic melanoma: An effective first-line test for the work-up of sentinel lymph node specimen. J Cutan Pathol. (2019) 10.1111/cup.13412, IHC; Human Read more (PubMed: 30632191) J Reichmann et al. Dual spindle formation in zygotes keeps parental genomes apart in early mammalian embryos, bioRxiv (2017) 10.1101/198275. IF; Mouse. Read more |
| . | |
 | ELISA of single stranded DNA using rabbit monoclonal anti-5-hmC (clone RM236) antibody. 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. |
 | Dot blot of double stranded DNA using rabbit monoclonal anti-5-hmC (clone 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. The pre-spotted membrane was then blotted with RM236. |
 | Immunohistochemical staining of formalin fixed and paraffin embedded human brain tissue sections, using rabbit monoclonal anti-5-hmC (clone RM236) antibody. |
 | Immunocytochemical staining of HeLa cells using 0.5 µg/mL anti-5-hmC antibody (RM236) (red). Actin filaments was labeled with fluorescein phalloidin (green). HeLa cells were fixed with 4% parafor-maldehyde and permeabilized with methanol (−20°C) before treatment with 2 N HCl for 30 min at 37°C to denature DNA. |
 | Direct ELISA of mouse brain genomic DNA using anti-5-hmC antibody, Clone 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. |
 | 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 1 µg 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. |
![Flow Cytometry analysis of 5-hmC expression in HEK293 cells using Anti-5-mC rabbit monoclonal antibody [RM236].The cells were fixed with ice-cold MeOH, permeabilizedwith 0.5% Triton X-100, denatured with 2N HCl, then stained with RM236 (anti-5-hmC, Blue) or with a negative control antibody(RM105, Red).](https://www.revmab.com/wp-content/uploads/2018/04/5hmc-MeOH.jpg) | Flow Cytometry analysis of 5-hmC expression in HEK293 cells using Anti-5-hmC rabbit monoclonal antibody, Clone RM236. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with RM236 (anti-5-hmC, Blue) or with a negative control antibody, Clone RM105, Red. |
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