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![Immunohistochemistry-Paraffin: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Paraffin-embedded rat brain. 5-Methylcytosine / 5-mC antibody diluted at 1:200.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Immunohistochemistry-Paraffin-NBP2-42814-img0008.jpg "Immunohistochemistry-Paraffin: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Paraffin-embedded rat brain. 5-Methylcytosine / 5-mC antibody diluted at 1:200.")
![Immunohistochemistry-Paraffin: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of human ovarian carcinoma. 5-Methylcytosine / 5-mC antibody [4111] dilution: 1:200.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Immunohistochemistry-Paraffin-NBP2-42814-img0002.jpg)
![Immunohistochemistry-Paraffin: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of mouse testis. 5-Methylcytosine / 5-mC antibody [4111] dilution: 1:200.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Immunohistochemistry-Paraffin-NBP2-42814-img0003.jpg)
![Immunohistochemistry-Paraffin: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of mouse urinary bladder. 5-Methylcytosine / 5-mC antibody [4111] dilution: 1:200.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Immunohistochemistry-Paraffin-NBP2-42814-img0004.jpg)
![Immunohistochemistry Whole-Mount: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of 24 hours-post-fertilization zebrafish embryo. 5-Methylcytosine / 5-mC antibody [4111] dilution: 1:50.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Immunohistochemistry-Whole-Mount-NBP2-42814-img0005.jpg)
![Methylated DNA Immunoprecipitation: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of 24 hours-post-fertilization zebrafish embryo. 5-Methylcytosine / 5-mC antibody [4111] dilution: 1:50.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Methylated-DNA-Immunoprecipitation-NBP2-42814-img0006.jpg)
![Dot Blot: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of anti-5-mC antibody with the synthetic DNA controls . DNA samples (0.5 to 50 ng) were spotted onto positively charged nylon membrane and blotted with 5-mC antibody at a dilution of 1:500. A: Unmethylated DNA fragment B: DNA fragment containing 5-methylcytosine C: DNA fragment containing 5-hydroxymethylcytosine D: DNA fragment containing 5-formylcytosine E: DNA fragment containing 5-carboxylcytosine .](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Dot-Blot-NBP2-42814-img0001.jpg)
![Methylated DNA Immunoprecipitation: 5-MethylCytosine Antibody (4111) [NBP2-42814] - Analysis of human genomic DNA (500 ng) mixed with methylated cytosine standard kit, using 5-Methylcytosine/ 5-mC antibody at 1:500 dilution. Analysized with semi-quantitative PCR and Immunprecipitated DNA was plotted as % of input DNA.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-4111-Methylated-DNA-Immunoprecipitation-NBP2-42814-img0007.jpg)
![Immunocytochemistry/Immunofluorescence: 5-hmC Antibody (RM236) [NBP2-61469] - Immunocytochemical staining of HeLa cells using NBP2-61469 at 0.5 ug/ml (red) . Actin filaments was labeled with fluorescein phalloidin green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (-20C) before treatment with 2 N HCl for 30 min at 37C to denature the DNA.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-Immunocytochemistry-Immunofluorescence-NBP2-61469-img0010.jpg "Immunocytochemistry/Immunofluorescence: 5-hmC Antibody (RM236) [NBP2-61469] - Immunocytochemical staining of HeLa cells using NBP2-61469 at 0.5 ug/ml (red) . Actin filaments was labeled with fluorescein phalloidin green). HeLa cells were fixed with 4% paraformaldehyde and permeabilized with methanol (-20C) before treatment with 2 N HCl for 30 min at 37C to denature the DNA.")
![Immunohistochemistry: 5-hmC Antibody (RM236) [NBP2-61469] - Immunohistochemical staining of formalin fixed and paraffin embedded Human brain tissue sections, using NBP2-61469](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-Immunohistochemistry-NBP2-61469-img0009.jpg)
![Flow Cytometry: 5-hmC Antibody (RM236) [NBP2-61469] - Flow Cytometry analysis of 5-hmC expression in HEK293 cells using NBP2-61470. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with NBP2-61469 (Blue) or with a negative control antibody (red)](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-Flow-Cytometry-NBP2-61469-img0014.jpg)
![ELISA: 5-hmC Antibody (RM236) [NBP2-61469] - Direct ELISA of Mouse brain genomic DNA using Anti-5-hmC antibody (NBP2-61469). The plate was directly coated with different concentrations of genomic DNA isolated from Mouse brain tissue. NBP2-61469 was used as the primary antibody, and a HRP conjugated Anti-Rabbit IgG as the secondary antibody.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-ELISA-NBP2-61469-img0002.jpg)
![ELISA: 5-hmC Antibody (RM236) [NBP2-61469] - ELISA of single stranded DNA using Anti-5-hmC antibody (NBP2-61469). 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 NBP2-61469 was used as the primary antibody, and an alkaline phosphatase conjugated Anti-Rabbit IgG as the secondary antibody.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-ELISA-NBP2-61469-img0011.jpg)
![ELISA: 5-hmC Antibody (RM236) [NBP2-61469] - Direct ELISA of Mouse brain genomic DNA using Anti-5-hmC antibody (NBP2-61469). The plate was directly coated with different concentrations of genomic DNA isolated from Mouse brain tissue. NBP2-61469 was used as the primary antibody, and a HRP conjugated Anti-Rabbit IgG as the secondary antibody.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-ELISA-NBP2-61469-img0012.jpg)
![Dot Blot: 5-hmC Antibody (RM236) [NBP2-61469] - Dot blot of double stranded DNA using NBP2-61469. The membrane was pre-spotted with double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-Dot-Blot-NBP2-61469-img0013.jpg)
![Methylated DNA Immunoprecipitation: 5-hmC Antibody (RM236) [NBP2-61469] - hMeDIP was performed using Anti-5-hmC antibody (NBP2-61469) 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.](http://images.novusbio.com/fullsize/5-hmC-Antibody-RM236-Methylated-DNA-Immunoprecipitation-NBP2-61469-img0015.jpg)
![Immunocytochemistry/Immunofluorescence: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - Immunocytochemical staining of HeLa cells using NBP2-61470 (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-Immunocytochemistry-Immunofluorescence-NBP2-61470-img0003.jpg "Immunocytochemistry/Immunofluorescence: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - Immunocytochemical staining of HeLa cells using NBP2-61470 (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).")
![Flow Cytometry: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - Flow Cytometry analysis of 5-MethylCytosine expression in HEK293 cells using NBP2-61470. The cells were fixed with ice-cold MeOH, permeabilized with 0.5% Triton X-100, denatured with 2N HCl, then stained with NBP2-61470 (Blue) or with a negative control antibody](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-Flow-Cytometry-NBP2-61470-img0008.jpg)
![ELISA: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - ELISA of single stranded DNA using NBP2-61470. 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 NBP2-61470 was used as the primary antibody, and an alkaline phosphatase conjugated anti-Rabbit IgG as the secondary antibody.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-ELISA-NBP2-61470-img0001.jpg)
![ELISA: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - Direct ELISA of HeLa cell genomic DNA using NBP2-61470. The plate was directly coated with different concentrations of genomic DNA isolated from HeLa cells. NBP2-61470 was used as the primary antibody, and a HRP conjugated Anti-Rabbit IgG as the secondary antibody.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-ELISA-NBP2-61470-img0006.jpg)
![Dot Blot: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - Dot blot of double stranded DNA using rabbit monoclonal NBP2-61470 antibody. The membrane was pre-spotted with double stranded 5-Hydroxymethylcytosine 5-hmC) DNA, 5-Methylcytosine 5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with NBP2-61470](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-Dot-Blot-NBP2-61470-img0007.jpg)
![Methylated DNA Immunoprecipitation: 5-MethylCytosine Antibody (RM231) [NBP2-61470] - MeDIP was performed using NBP2-61470 at a 2:1 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.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-RM231-Methylated-DNA-Immunoprecipitation-NBP2-61470-img0009.jpg)
![Immunocytochemistry/Immunofluorescence: 5-MethylCytosine Antibody (33D3) [NBP2-54609] - HeLa cells were stained with the antibody against 5-mC and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-33D3-Immunocytochemistry-Immunofluorescence-NBP2-54609-img0022.jpg "Immunocytochemistry/Immunofluorescence: 5-MethylCytosine Antibody (33D3) [NBP2-54609] - HeLa cells were stained with the antibody against 5-mC and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.")
![Surface Plasmon Resonance: 5-MethylCytosine Antibody (33D3) [NBP2-54609] - A synthesized biotin-labeled 5-mC conjugate was immobilized on a sensorchip. Briefly, two flowcells were prepared by sequential injections of EDC/NHS, streptavidin, and ethanolamine. One of these flowcells served as negative control, while biotinylated 5-mC conjugate was injected in the other one, to get an immobilization level of 55 response units (RU). All SPR experiments were performed, using HBS-N buffer (10 mM HEPES,150 mM NaCl, pH 7.4), at a flow rate of 5 uL/min. Interaction assays involved injections of 2 different dilutions of the 5-mC monoclonal antibody over the biotinylated 5-mC conjugate and negative control surfaces, followed by a 3 minute washing step with HBS-N buffer. At the end of each cycle, the streptavidin surface was regenerated by injection of 0.1M citric acid (pH 3). The value of the dissociation constant (kd) obtained by global fitting and 1:1 Langmuir model is 65 nM.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-33D3-Ovalbumine-Surface-Plasmon-Resonance-NBP2-54609-img0015.jpg)
![Dot Blot: 5-MethylCytosine Antibody (33D3) [NBP2-54609] - To demonstrate the specificity of the antibody against 5-mC, a Dot blot analysis was performed using hmC, mC and C controls. 100 to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane. The antibody was used at a dilution of 1:300. Figure shows a high specificity of the antibody for the methylated control.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-33D3-Dot-Blot-NBP2-54609-img0018.jpg)
![Methylated DNA Immunoprecipitation: 5-MethylCytosine Antibody (33D3) [NBP2-54609] - Analysis was performed on 1 ug fragmented human genomic DNA using 0.2 ug of the monoclonal antibody against 5-mC. The fragmented DNA was spiked with controls (methylated DNA (meDNA) as a positive and unmethylated DNA (unDNA) as a negative control) prior to performing the IP. QPCR was performed with primer sets specific for the methylated and unmethylated DNA controls, and for a known methylated (TSH2B) and unmethylated (GAPDH) genomic region. The figure shows the recovery expressed as a percent of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-33D3-Methylated-DNA-Immunoprecipitation-NBP2-54609-img0023.jpg)
![Immunofluorescence: 5-MethylCytosine Antibody (C.15200006) [NBP2-59170] - HeLa cells were stained with the antibody against 5-mC and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (left) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-C-15200006-Immunofluorescence-NBP2-59170-img0002.jpg "Immunofluorescence: 5-MethylCytosine Antibody (C.15200006) [NBP2-59170] - HeLa cells were stained with the antibody against 5-mC and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (left) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.")
![Methylated DNA Immunoprecipitation: 5-MethylCytosine Antibody (C.15200006) [NBP2-59170] - Methylated DNA immunoprecipitation was performed on 1 ug fragmented human genomic DNA using 0.2 ug of the antibody against 5-mC and a MeDIP Kit. The fragmented DNA was spiked with the internal controls present in the kit (methylated DNA (meDNA) as a positive and unmethylated DNA (unDNA) as a negative control) prior to performing the IP. QPCR was performed with optimized primer sets, specific for the methylated and unmethylated DNA controls, and for a known methylated (TSH2B) and unmethylated (GAPDH) genomic region. Figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/5-MethylCytosine-Antibody-C-15200006-Methylated-DNA-Immunoprecipitation-NBP2-59170-img0001.jpg)
![Methylated DNA Immunoprecipitation: 5-hmC Antibody (C.15220001) [NBP2-59178] - Hydroxymethylated DNA IP (hMeDIP) assays were performed using the antibody against 5-hydroxymethylcytosine, 5-hmC, 5-mC and cytosine DNA standards and Rat IgG. DNA fragments are 300-500 bp. 1 ug of mouse ES cells DNA was spiked with 0.025 ng of each DNA standard. The IP'd material has been analyzed by qPCR using the primer pairs specific to the control sequences. The obtained results are as follows: - hMeDIP on unmethylated control, with Rat IgG as negative control (0.06%, almost no recovery), with 5-hmC antibody (0.61%, almost no recovery) - hMeDIP on methylated control, with Rat IgG as negative control (0.03%, almost no recovery), with 5-hmC antibody (0.62%, almost no recovery) - hMeDIP on hydroxymethylated control, with Rat IgG as negative control (0.04%, almost no recovery), with 5-hmC (97.60% recovery, almost full recovery).](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-C-15220001-Methylated-DNA-Immunoprecipitation-NBP2-59178-img0002.jpg "Methylated DNA Immunoprecipitation: 5-hmC Antibody (C.15220001) [NBP2-59178] - Hydroxymethylated DNA IP (hMeDIP) assays were performed using the antibody against 5-hydroxymethylcytosine, 5-hmC, 5-mC and cytosine DNA standards and Rat IgG. DNA fragments are 300-500 bp. 1 ug of mouse ES cells DNA was spiked with 0.025 ng of each DNA standard. The IP'd material has been analyzed by qPCR using the primer pairs specific to the control sequences. The obtained results are as follows: - hMeDIP on unmethylated control, with Rat IgG as negative control (0.06%, almost no recovery), with 5-hmC antibody (0.61%, almost no recovery) - hMeDIP on methylated control, with Rat IgG as negative control (0.03%, almost no recovery), with 5-hmC antibody (0.62%, almost no recovery) - hMeDIP on hydroxymethylated control, with Rat IgG as negative control (0.04%, almost no recovery), with 5-hmC (97.60% recovery, almost full recovery).")
![ELISA: 5-hmC Antibody (C.15220001) [NBP2-59178] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against 5-hmC in antigen coated wells. The antigen used was a 5-hmC base coupled to KHL. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:25,000.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-C-15220001-ELISA-NBP2-59178-img0001.jpg)
![Dot Blot: 5-hmC Antibody (C.15220001) [NBP2-59178] - (A): Approximately 200 ng, equivalent 10 pmol of C-bases, of the hmC (1), mC (2) and C (3) PCR controls were spotted on a membrane (Amersham Hybond-N+). The membrane was incubated with 5-hmC Antibody (C.15220001) followed by an HRP conjugated anti-rat secondary antibody. The membrane was exposed for 30 seconds.](http://images.novusbio.com/fullsize/5-hmC-Antibody-C-15220001-Dot-Blot-NBP2-59178-img0004.jpg)
![Dot Blot: 5-hmC Antibody (C.15220001) [NBP2-59178] - 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-base) of the hmC (1), mC (2) and C (3) PCR controls were spotted on a membrane (Amersham Hybond-N+). The membrane was incubated with 4 ug/ml (dilution 1:500) of the 5-hmC Antibody (C.15220001), followed by an HRP conjugated anti-rat secondary antibody. The membrane was exposed for 30 seconds.](http://images.novusbio.com/fullsize/5-hmC-Antibody-C-15220001-Dot-Blot-NBP2-59178-img0005.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.](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.")
![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)
![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)
![Dot Blot: 5-hmC Antibody (C.15200200) [NBP2-59177] - 5-Hydroxymethylcytosine Antibody (C.15200200) [NBP2-59177] - Dot Blot analysis of the antibody with the C, mC and hmC PCR controls 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-bases) of the hmC (1), mC (2) and C (3) PCR controls from the antibody (dilution 1:500). The membranes were exposed for 30 seconds.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-C-15200200-Dot-Blot-NBP2-59177-img0001.jpg "Dot Blot: 5-hmC Antibody (C.15200200) [NBP2-59177] - 5-Hydroxymethylcytosine Antibody (C.15200200) [NBP2-59177] - Dot Blot analysis of the antibody with the C, mC and hmC PCR controls 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-bases) of the hmC (1), mC (2) and C (3) PCR controls from the antibody (dilution 1:500). The membranes were exposed for 30 seconds.")
![Methylated DNA Immunoprecipitation: 5-hmC Antibody (C.15200200) [NBP2-59177] - 5-Hydroxymethylcytosine Antibody (C.15200200) [NBP2-59177] - An hydroxymethylated DNA IP (hMeDIP) was performed using the antibody directed against 5-hydroxymethylcytosine. The IgG isotype antibodies from mouse was used as negative control. The DNA was prepared with the GenDNA module of the hMeDIP kit and sonicated to have DNA fragments of 300-500 bp. 1 ug of human HeLa cells DNA were spiked with non-methylated, methylated, and hydroxymethylated PCR fragments. The IP'd material has been analysed by qPCR using the primer pair specific for the 3 different control sequences. The obtained results show that the mouse monoclonal for 5-hmC is highly specific for this base modification (no IP with non-methylated or methylated C bases containing fragments).](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-C-15200200-Methylated-DNA-Immunoprecipitation-NBP2-59177-img0002.jpg)
![ELISA: 5-hmC Antibody (C.15200200) [NBP2-59177] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against 5-hmC in antigen coated wells. The antigen used was KHL coupled to 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.](http://images.novusbio.com/fullsize/5-hmC-Antibody-C-15200200-ELISA-NBP2-59177-img0004.jpg)
![Dot Blot: 5-hmC Antibody (513) [NBP2-43568] - Analysis of anti- 5-Hydroxymethylcytosine / 5-hmC antibody [513] with DNA samples DNA samples (50 to 500 ng as indicated) were spotted onto positively charged nylon membrane and blotted with 5-Hydroxymethylcytosine / 5-hmC antibody [513] at a dilution of 1:500. A: Genomic DNA derived from 293T cells. B: Genomic DNA derived from human TET-1 transfected 293T cells.](http://images.novusbio.com/fullsize/5-hmC-Antibody-513-Dot-Blot-NBP2-43568-img0001.jpg "Dot Blot: 5-hmC Antibody (513) [NBP2-43568] - Analysis of anti- 5-Hydroxymethylcytosine / 5-hmC antibody [513] with DNA samples DNA samples (50 to 500 ng as indicated) were spotted onto positively charged nylon membrane and blotted with 5-Hydroxymethylcytosine / 5-hmC antibody [513] at a dilution of 1:500. A: Genomic DNA derived from 293T cells. B: Genomic DNA derived from human TET-1 transfected 293T cells.")
![Dot Blot: 5-hmC Antibody (513) [NBP2-43568] - Analysis of anti-5-hmC antibody with the synthetic DNA controls . DNA samples (0.5 to 50 ng as indicated) were spotted onto the positively charged Nylon membrane and blotted with 5-hmC antibody at a dilution of 1:500. A: Unmethylated DNA fragment B: DNA fragment containing 5-methylcytosine C: DNA fragment containing 5-hydroxymethylcytosine D: DNA fragment containing 5-formylcytosine E: DNA fragment containing 5-carboxylcytosine.](http://images.novusbio.com/fullsize/5-hmC-Antibody-513-Dot-Blot-NBP2-43568-img0002.jpg)
![Methylated DNA Immunoprecipitation: 5-hmC Antibody (513) [NBP2-43568] - Analysis of Human genomic DNA (500 ng) mixed with methylated cytosine standard kit, using 5-Hydroxymethylcytosine/ 5-hmC antibody [513] at 1:500 dilution. Analysized with semi-quantitative PCR and Immunprecipitated DNA was plotted as % of input DNA.](http://images.novusbio.com/fullsize/5-hmC-Antibody-513-Methylated-DNA-Immunoprecipitation-NBP2-43568-img0004.jpg)
![Dot Blot: 5-hmC Antibody [NBP2-43569] - Analysis of anti-5-hmC antibody with the synthetic DNA controls . DNA samples (0.5 to 50 ng as indicated) were spotted onto the positively charged Nylon membrane and blotted with 5-hmC antibody at a dilution of 1:50000. A: Unmethylated DNA B: DNA containing 5-methylcytosine C: DNA containing 5-hydroxymethylcytosine D: DNA containing 5-formylcytosine E: DNA containing 5-carboxylcytosine.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-Dot-Blot-NBP2-43569-img0005.jpg "Dot Blot: 5-hmC Antibody [NBP2-43569] - Analysis of anti-5-hmC antibody with the synthetic DNA controls . DNA samples (0.5 to 50 ng as indicated) were spotted onto the positively charged Nylon membrane and blotted with 5-hmC antibody at a dilution of 1:50000. A: Unmethylated DNA B: DNA containing 5-methylcytosine C: DNA containing 5-hydroxymethylcytosine D: DNA containing 5-formylcytosine E: DNA containing 5-carboxylcytosine.")
![Methylated DNA Immunoprecipitation: 5-hmC Antibody [NBP2-43569] - Human genomic DNA (150 ng) mixed with methylated cytosine standard kit were subjected to MeDIP with 5-Hydroxymethylcytosine/ 5-hmC antibody at dilution of 1:2000 and analyzed with semi-quantitative PCR. The immunoprecipitated DNA was plotted as % of input DNA.](http://images.novusbio.com/fullsize/5-Hydroxymethylcytosine-Antibody-Methylated-DNA-Immunoprecipitation-NBP2-43569-img0004.jpg)