Cat. # | Size | Qty. | Price |
---|---|---|---|
32205S | 100 µl |
|
REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 39 |
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
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro241 of human ARH3 protein.
ADP-ribosylation is a reversible post-translational modification (PTM) where a single unit or polymers of ADP-ribose (ADPr) groups from β-nicotinamide adenine dinucleotide (NAD+) are transferred onto specific residues of target proteins (1). In addition to proteins, DNA and RNA molecules have also been identified as targets of ADP-ribosylation (2,3). This modification is catalyzed by a diverse group of ADP-ribosyltransferases (ARTs), with the majority of them functioning to add a single ADPr unit (MARylation) while the deposition of multiple ADPrs are done by poly (ADP-ribose) polymerases (PARPs). Reversal of ADP-ribosylation is accomplished by the evolutionarily distinct macrodomain and ADP-ribosylhydrolase (ARH) protein families (4). Specifically, poly (ADP-ribose) glycohydrolase (PARG) or ADP-ribosylhydrolase 3 (ARH3) degrade PARylation down to single ADPr units and MacroD1, MacroD2, TARG1, or ARH1 fully remove them from target residues (5). ADP-ribosylation and the enzymes that regulate it are involved in a wide range of cellular processes, such as DNA repair, chromatin regulation, transcription, and cellular stress response (1,4). Moreover, ADP-ribosylation has been shown to be a therapeutically critical PTM in various cancers, neurodegenerative diseases, and inflammation (6).
The ARH family of ADP-ribose hydrolases contains three proteins with similarities in amino acid sequence but differences in their enzymatic activities. ARH3, also known as ADPRHL2, is a ubiquitous protein that localizes to the nucleus, cytosol, and mitochondria in a cell-type specific manner (7-9). ARH3 exhibits partial redundancy with PARG in their preference for hydrolysis of DNA damage-induced serine ADP-ribosylation which underlies a key role for these enzymes in maintaining genomic stability. Arh3-KO mice have been shown to develop increased brain infarction in response to ischemia-reperfusion injury (10) and human ARH3-deficiency is a rare recessive autosomal disorder characterized by neurodegeneration and early death (11,12).
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