![Dot Blot: 5-hmC Antibody [NBP2-59184] - To demonstrate the specificity of the antibody against 5-hmC, a Dot blot analysis was performed using hmC, mC and C controls. One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:1,000. Figure shows a high specificity of the antibody for the hydroxymethylated control.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-Dot-Blot-NBP2-59184-img0001.jpg "Dot Blot: 5-hmC Antibody [NBP2-59184] - To demonstrate the specificity of the antibody against 5-hmC, a Dot blot analysis was performed using hmC, mC and C controls. One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:1,000. Figure shows a high specificity of the antibody for the hydroxymethylated control.")
| Reactivity | Hu, Mu, Xp, ZeSpecies Glossary |
| Applications | DB, ELISA, IP |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Immunogen | The exact immunogen is propietary information. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Purity | Protein G purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Storage | Store at -20C. Avoid freeze-thaw cycles. |
| Buffer | PBS |
| Preservative | 0.05% Sodium Azide and 0.05% ProClin 300 |
| Purity | Protein G purified |
Secondary Antibodies |
Isotype Controls |
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![Methylated DNA Immunoprecipitation: 5-hmC Antibody [NBP2-59184] - hMeDIP (hydroxymethylated DNA IP) was performed using the antibody against 5-hydroxymethylcytosine. DNA from mouse ES cells was prepared and sonicated to obtain DNA fragments of 300-500 bp. One ug of sheared DNA was spiked with the unmethylated (C) methylated (mC), and hydroxymethylated (hmC) controls. hMeDIP was performed with 3.5 ug of the rabbit 5-hmC antibody and the IP'd DNA was analysed by qPCR using primers specific for the 3 different control sequences. Figure shows that the rabbit polyclonal antibody against 5-hmC is highly specific for the 5-hmC base modification (no IP with non-methylated or methylated C control fragments).](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-Methylated-DNA-Immunoprecipitation-NBP2-59184-img0003.jpg "Methylated DNA Immunoprecipitation: 5-hmC Antibody [NBP2-59184] - hMeDIP (hydroxymethylated DNA IP) was performed using the antibody against 5-hydroxymethylcytosine. DNA from mouse ES cells was prepared and sonicated to obtain DNA fragments of 300-500 bp. One ug of sheared DNA was spiked with the unmethylated (C) methylated (mC), and hydroxymethylated (hmC) controls. hMeDIP was performed with 3.5 ug of the rabbit 5-hmC antibody and the IP'd DNA was analysed by qPCR using primers specific for the 3 different control sequences. Figure shows that the rabbit polyclonal antibody against 5-hmC is highly specific for the 5-hmC base modification (no IP with non-methylated or methylated C control fragments).")
![ELISA: 5-hmC Antibody [NBP2-59184] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against 5-hmC, crude serum and flow through, in antigen coated wells. The antigen used was the 5-hmC base coupled to BSA. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:2,800.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-ELISA-NBP2-59184-img0002.jpg "ELISA: 5-hmC Antibody [NBP2-59184] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against 5-hmC, crude serum and flow through, in antigen coated wells. The antigen used was the 5-hmC base coupled to BSA. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:2,800.")
![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")