Imprint® Monoclonal Anti-5-hydroxymethylcytosine antibody produced in rat
| Name | Imprint® Monoclonal Anti-5-hydroxymethylcytosine antibody produced in rat |
|---|---|
| Supplier | Sigma-Aldrich |
| Catalog | SAB4800002 |
| Prices | $885.00 |
| Sizes | 100 µg |
| Host | Rat |
| Applications | MeDIP ELISA |
| Species Reactivities | Mouse, Human |
| Description | Rat |
| Conjugate | unconjugated |
| Supplier Page | Shop |
Product images
ImmunoblottingAnti-5-hydroxymethylcytosine: Cat. No. SAB4800002: Dot blot analysis of 5-hydroxymethylcytosine and 5-methylcytosine monoclonal antibodies with cytosine, C, methylcytosine, mC, and hydroxymethylcytosine, hmC, PCR controls.
Figure A: Approximately 200 ng, equivalent 10 pmol of C-base, of hmC, 1, mC, 2, and C, 3, PCR controls were spotted on a membrane. The membrane was incubated with 4 μg/mL 5-hmC rat monoclonal antibody, followed by an HRP conjugated secondary antibody. The membrane was exposed for 30 seconds.
Figure B: Incubation of the same membrane with the 5-methylcytosine Mouse monoclonal antibody, Cat. No. SAB4800001, diluted 1:250. The membrane was not stripped after the 5-hmC incubation. The left spot represents the remaining hmC signal, which confirms that an equal amount of mC bases was spotted at position 2.
ImmunoblottingAnti-5-hydroxymethylcytosine: Cat. No. SAB4800002: Dot blot analysis of the 5-hydroxymethylcytosine rat monoclonal antibody with cytosine, C, methylcytosine, mC, and hydroxymethylcytosine, hmC, PCR controls. 200 to 2 ng, equivalent of 10 to 0.1 pmol of C-base, of hmC, 1, mC, 2, and C, 3, PCR controls were spotted on a membrane. The membrane was incubated with 4 μg/mL, 1:500 dilution, of the antibody, followed by an HRP conjugated secondary antibody. The membrane was exposed for 30 seconds.
ChIP Results ChartAnti-5-hydroxymethylcytosine: Cat. No. SAB4800002: An hydroxymethylated DNA IP, hMeDIP, was performed using the Anti-5-hydroxymethylcytosine antibody and DNA from Mouse ES cells. 1 μg of Mouse ES cells DNA was spiked with 0.025 ng of positive or negative DNA standards. IP minutesd material was analyzed by qPCR using the primer pairs specific to the control sequences.
Columns, left to right,
1. Rat IgG negative control, Unmethylated Control, IgG,
2. 5-hmC antibody negative control, Unmethylated Control, 5-hmC,
3. Rat IgG negative control, Methylated Control, IgG,
4. 5-hmC antibody negative control, Methylated Control, 5-hmC,
5. Rat IgG negative control, Hydroxymethylated Control, IgG, 6. 5-hmC antibody, Hydroxymethylated Control, 5-hmC.
ELISAAnti-5-hydroxymethylcytosine: Cat. No. SAB4800002: Determination of the 5-hydroxymethylcytosine, 5-hmC, rat monoclonal antibody titer. An ELISA was performed using a serial dilution of the antibody 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.