Western blot analysis of extracts from various cell lines using HPF1 Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
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: 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
HPF1 Antibody recognizes endogenous levels of total HPF1 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu241 of human HPF1 protein. Antibodies are purified by peptide affinity chromatography.
Poly (ADP-ribose) polymerases (PARPs) are a family of enzymes that catalyze ADP-ribosylation (ADPr), a reversible posttranslational modification of proteins that regulates various cellular processes. PARP-1 and PARP-2 are two well-characterized family members that catalyze ADPr on a wide range of substrates in response to DNA damage (1). Histone PARylation factor 1 (HPF1), also referred to as C4orf27, is an accessory protein to PARP1/2 that directly binds to these enzymes and alters their ADPr amino acid specificity from aspartate/glutamate to serine. (2). HPF1 is specifically required to carry out ADPr modifications on serine residues for both histone and non-histone substrates, as serine ADPr does not occur in cells lacking HPF1 (3). This is critically important, as serine has been shown to be the most abundant ADPr residue in cells affected by DNA damage (4). Mechanistically, PARP1/2 have been described as incomplete enzymes, and crystal structure analysis shows that HPF1-PARP1/2 binding confers a joint active site, which may explain the switch from aspartate/glutamate specificity to serine (2). Additionally, HPF1 knockout cells are more sensitive to PARP inhibitors, suggesting that drug manufacturers will need to take HPF1 binding into account when developing effective inhibitors for these enzymes (1).
Explore pathways + proteins related to this product.
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