| Lot | A17-0042 |
|---|---|
| Concentration | 0.7 µg/µl |
| Species reactivity | Human, mouse |
| Type | Polyclonal |
| Purity | Affinity purified |
| Host | Rabbit |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP * | 1.5 μg/ChIP | Fig 1 |
| ELISA | 1:1,000 | Fig 2 |
| Western Blotting | 1:500 | Fig 3 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
Figure 1 ChIP results obtained with the Diagenode antibody directed against MBD1
ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against MBD1 (Cat. No. pAb-078-050) and optimized PCR primer sets. Sheared chromatin from 1x10e6 cells and 1.5 μg of antibody were used per ChIP experiment. Beads only were used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the MLH1 gene (used as a positive control) and CDC6 gene (used as a negative control). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2 Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human MBD1 (Cat. No. pAb-078-050), crude serum and Flow Through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the purified antibody was estimated to be 1:20,000.
Figure 3 Western blot analysis using the Diagenode antibody directed against MBD1
Nuclear extracts of HeLa cells (40 μg) were analysed by Western blot using the Diagenode antibody against MBD1 (Cat. No. pAb-078-050) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.
MBD1 (UniProt/Swiss-Prot entry Q9UIS9) is a transcriptional repressor that specifically binds to methylated CpG dinucleotides in promoter sequences. MBD1 acts by recruiting a variety of histone deacetylases (HDAC’s) and chromatin remodelling factors. MBD1-dependent transcriptional repression is mediated by ATF7IP through the recruitment of factors such as the histone methyltransferase SETDB1. MBD1 probably forms a complex with SETDB1 and ATF7IP which couples DNA methylation to H3K9 trimethylation and represses transcription.
| ELISA Enzyme-linked immunosorbent assay. Read more |
| WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more |
| ChIP-qPCR (ab) Read more |
| Datasheet MBD1 C15410078 DATASHEET Datasheet description | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
How to properly cite our product/service in your workWe strongly recommend using this: MBD1 Antibody (Hologic Diagenode Cat# C15410078 Lot# A17-0042). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
Novel role of UHRF1 in the epigenetic repression of the latent HIV-1. |
H19 lncRNA controls gene expression of the Imprinted Gene Network by recruiting MBD1. |