Epigenetic studies often use antibodies for immunoprecipitation, Western blot or immunolocalization experiments to analyze modified nucleic acids, transcription cofactor proteins and histones. DNA immunoprecipitation based experiments are often followed by microarray or next generation high-throughput sequencing.
Me-DIP enriches for methylated or hydroxymethylated DNA, and can be carried out in the presence of cofactors such as heterodimer partners, competitors or small molecule binding inhibitors (1).
Chromatin immunoprecipitation (ChIP), which uses modification specific histone antibodies, is the most commonly used method to analyze histone modifications associated with genes (2). The isolated DNA may be used for next-generation sequencing for genome wide studies.
Antibodies against chromatin modifying enzymes such as histone acetyltransferases/deacetylases, histone methyltransferases/demethylases, histone kinases/phosphatases, and ubiquitin ligases can be used to probe their functions and locations on chromatin.
RNA binding protein immmunoprecipitation (RIP) uses antibodies to pull down RNA binding proteins, which co-precipitate with their interacting RNA molecules for downstream analysis by RT-PCR (3) followed by sequencing.
FAQs for Antibodies
- What dilution should be used with Dnmt1 Amino-terminal Ab?
- What is the species cross reactivity of Dnmt1 Amino-terminal Ab?
- Can Dnmt1 Amino-terminal Ab be used to detect mouse and rat?
- How much of NEB Dnmt1 should be used as a control when using Dnmt1 Amino-terminal Ab?
- How much protein should be run on the gel for detection with Dnmt1 Amino-terminal Ab?
Protocols for Antibodies
- Cross-linking of IgG to Protein A or G Beads
- Goat Anti-Mouse IgG Magnetic Beads Protocol
- Goat Anti-Rabbit IgG Magnetic Beads Protocol
- Goat Anti-Rat IgG Magnetic Beads (S1433)
- Immunoprecipitation using Protein A/G Magnetic Beads
- Phage Display: Solution-phase Panning with Affinity Bead Capture
- Protocol for Anti-SNAP-tag® Antibody (Polyclonal) (P9310)
- Purification of IgG using Protein A/G Magnetic Beads
If all cells are created from the same genetic material, why are there so many different cell types? Listen to Sriharsa Pradhan, Senior Scientist, RNA Biology at NEB, as he describes how DNA is methylated and how this affects the path of reading the DNA code the same way an obstruction would derail a train off its tracks.