Western blot analysis of extracts from Mv1Lu cells treated with UV or hydroxyurea (HU) for the indicated times, using Phospho-p95/NBS1 (Ser343) Antibody.
|REACTIVITY||H M Mi|
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
Phospho-p95/NBS1 (Ser343) Antibody detects endogenous levels of p95/NBS1 only when phosphorylated at serine 343.
Human, Mouse, Mink
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser343 of human p95/NBS1. Antibodies are purified by protein A and peptide affinity chromatography.
Nijmegen breakage syndrome (NBS) is characterized by growth retardation, mental disability, immunodeficiency, defects in cell cycle checkpoints, an increased propensity for cancer, and sensitivity to ionizing radiation (1). Repair of radiation-induced DNA double-strand breaks is dependent on the multifunctional MRN complex containing Mre11, Rad50, and the NBS1 gene product p95/NBS1 (also called p95 or nibrin) (2). p95/NBS1 is a protein with a forkhead-associated domain and a BRCT repeat that regulate interaction with MDC1 and are essential for proper G2/M DNA-damage checkpoint function (3). NBS1 is critical for homologous recombination following DNA double strand breaks. This activity requires CDK-dependent association with CtIP and subsequent phosphorylation by ATM (4). ATM interacts with and phosphorylates p95/NBS1 at Ser278 and Ser343 after exposure to ionizing radiation (5,6).
Explore pathways + proteins related to this product.