Dnmt1-Trap Affinity Reagents

Novel Binding Protein for Dnmt1 Studies

Analyze Dnmt1 proteins in a tube

DNA methylation is the post-replicative transfer of a methyl group to the C5 position of a cytosine base. It was the first epigenetic modification identified and has been intensively studied for more than half a century. By now it is clear that Dnmt1, the major eukaryotic DNA methyltransferase, faithfully maintains genome-wide methylation patterns and plays an essential role in the epigenetic network controlling gene expression and genome stability during development. However, the molecular mechanisms that ultimately control DNA methylation still remain elusive. Use Dnmt1-Trap for very fast, efficient and specific pull-down of endogenous Dnmt1 from cell extracts. A single step protocol yields a much cleaner result, free from contaminants, in under 30 minutes. One can also use Dnmt1-Trap to co-immunoprecipitate interacting factors for identification and characterization (e.g. through mass spec).

 



 

LITERATURE
Protocols
FAQs
 
RELATED PRODUCTS
Other Nano-Trap Reagents
Dnmt1 Antibodies
Live Cell Imaging  Microscopes

 

Dnmt1-Traps utilize super-high affinity Camelidae antibody fragments that may be used for immuno-precipitation, immuno-purification and immuno-pull down experiments with up to 10-fold better purity and yield than that of classic mouse monoclonal antibodies. Compatible with a variety of source materials, Nano-Traps may be used with mammalian cells, tissues & organs, bacteria, yeast and even plants. This reagent allows Dnmt1 to be used for immunoprecipitations, Co-IP, mass spectroscopy, and enzyme activity measurements.

 

Nano-Traps

 

Dnmt1 Protein Gel

Dnmt1-Trap for immunoprecipitatations

The complementary rat monoclonal Dnmt1 antibody 2E8 is well-suited to detect a very precise band at 180 kDa in Western Blot analysis (see results below). Furthermore the Dnmt1 antibody 2E8 shows a distinct nuclear Dnmt1 immunofluorescence staining in G-phase cells, as well the characteristic replication pattern in S phase cells where maintenance methylation takes place. Used here for demonstrating the pristine immunoprecitation one can expect from any of our Nano-Trap products. In this case, Dnmt1 was pulled down from a protein extract of HEK 293T cells.

 

Input (I), non-bound (FT) and bound (B) fractions were separated by SDS-PAGE followed by Coomassie staining and Western Blotting (using antibody 2E8). Unlike conventional antibodies, the Dnmt1-Trap is free from heavy and/or light chain contaminants and therefore yields cleaner results.

 

 

 

 

 

 

 

 

 

Immunofluorescence

Dnmt1 Immunofluorescence

 

Shown to the left are HeLa cells fixed witg 3.7% PFA . Primary antibody: 1/100 Dnmt1 antibody 2E8 incubation for 1 h at RT. Secondary antibody: (top) 1/400 anti-rat-Alexa488 incubation for 1 h at RT, (bottom) 1/400 anti-rat-Alexa568 incubation for 1 h at RT. Characteristic punctate staining of Dnmt1 at replication foci in the nucleus of cells in S phase.

 

 

 

 

 

 

 

 

 

 

 

 

 

Camelidae Antibody

Camelidae single-domain antibodies are like IgGs on steroids

The family of animals known as Camelidae (camels, dromedaries, llamas and alpacas) produce functional antibodies devoid of light chains, so called "heavy chain" antibodies. These heavy chain antibodies recognize and bind their antigens via a single variable domain. When cleaved from their carboxy tail, these barrel-shaped structures (2x3 nm) are extraordinarily small, naturally-occurring, and intact antigen binding fragments (MW  of 13 kDa). These fragments, called Nanobodies, are characterized by high specificity and affinities in the low nanomolar range, and dissociation constants in the sub-nanomolar range (typically 10- to 100-fold better than mouse IgGs). The compact size of Nanobodies makes them extremely stable at temperatures up to 70°C, and functional even in 2M NaCl or 0.5% SDS. These small and powerful antibody fragments can be used in a variety of unique applications. They will open up your research possibilities.

 

 

 

 

SPECIFICATIONS

Configuration:

 

Dnmt1-TrapA

Specific Camelidae antibody linked to agarose bead

Dnmt1-Trap

Uncoupled and purified Camelidae antibody

Part Numbers DTA010, DTA020, DTA100, DTA200, DTA400, DTAK020, DT250

Particle Size:

 

Dnmt1-Trap A

~90 µm  when coupled to an agarose beads

Dnmt1-Trap

No particle coupled

Storage Buffer:  

Dnmt1-Trap A

20% EtOH

Dnmt1-Trap

1x PBS; Preservative: 0.01% Sodium Azide

Storage and Stability:

 

Dnmt1-Trap A

store at 4°C; stable for one year. Do not freeze.

Dnmt1-Trap

store at 4°C; stable for one year. Do not freeze.



Dnmt1 Traps

ORDERING - Dnmt1-Traps Coupled to Agarose

 

 

Binding Controls
Spin Columns for agarose beads