Cat. # | Size | Qty. | Price |
---|---|---|---|
99105S | 100 µl |
|
REACTIVITY | H M R Mk |
SENSITIVITY | Endogenous |
MW (kDa) | 58, 60 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human TDG protein.
Methylation of DNA at cytosine residues is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and mammalian development (1,2). 5-methylcytosine is a repressive epigenetic mark established de novo by two enzymes, DNMT3A and DNMT3B, and is maintained by DNMT1 (3,4). 5-methylcytosine was originally thought to be passively depleted during DNA replication. However, subsequent studies have shown that Ten-Eleven Translocation (TET) proteins TET1, TET2, and TET3 can catalyze the oxidation of methylated cytosine to 5-hydroxymethylcytosine (5-hmC) (5). Additionally, TET proteins can further oxidize 5-hmC to form 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC), both of which are excised by thymine-DNA glycosylase (TDG), effectively linking cytosine oxidation to the base excision repair pathway and supporting active cytosine demethylation (6,7). Knockout or catalytic inactivation of TDG leads to embryonic lethality, due in part to the loss of DNA methylation patterns at the promoters and enhancers of developmental genes (8,9). TDG is commonly SUMOylated at Lys330, although the exact consequence of this modification is not yet fully understood. SUMOylation has been reported to help TDG dissociate from its abasic product, thereby increasing catalytic turnover (10-12). Additional studies suggest that SUMOylation affects TDG’s cellular localization or lowers its base excision activity, allowing it to act as a ‘reader’ protein for 5-fC and 5-caC modified DNA (13-14).
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