Cat. # | Size | Qty. | Price |
---|---|---|---|
3522S | 100 µl |
|
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 59 |
SOURCE | Rabbit |
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
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser114 and Thr118 of human PNK1. Antibodies are purified by protein A and peptide affinity chromatography.
PNK (polynucleotide kinase) is a DNA repair enzyme that participates in single strand break repair and non-homologous end rejoining (NHEJ) for double strand breaks. PNK possesses a 5'-DNA kinase activity and a 3'-DNA phosphatase activity (1,2). It has three domains, a C-terminal kinase domain, a central phosphatase domain, and an N-terminal forkhead associated (FHA) domain that is responsible for protein-protein interactions. Reduction in expression of PNK by RNAi sensitizes cells to ionizing radiation and topoisomerase I inhibitors (3)
Phosphorylation of PNK1 at Ser114/ Thr118 was independently identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's MS/MS platform for phosphorylation site discovery. Phosphorylation of PNK1 at Ser114/ Thr118 was observed in extracts of various cell lines following UV treatment. For additional information please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org.
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