Cat. # | Size | Qty. | Price |
---|---|---|---|
9386T | 20 µl |
|
|
9386S | 100 µl |
|
REACTIVITY | All |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Mouse IgM |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Peptide ELISA (DELFIA) | 1:2000 |
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.
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: 262
This protocol is intended for immunoprecipitation of native proteins for western immunoblot analysis or kinase activity assays.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
10X Cell Lysis Buffer: (#9803) To prepare 10 ml of 1X cell lysis buffer, add 1 ml cell lysis buffer to 9 ml dH2O, mix.
NOTE: Add 1 mM PMSF (#8553) immediately prior to use.
posted January 2016
Protocol Id: 884
(DELFIA® is a registered trademark of PerkinElmer, Inc.)
posted June 2005
revised September 2007
Protocol Id: 34
All Species Expected
Monoclonal antibody is produced by immunizing animals with phospho-Thr-containing peptides.
Much of the dynamic behavior of cellular proteins, including the regulation of molecular interactions (1), subcellular localization (2), and transcriptional regulation (3) is controlled by a variety of post-translational modifications (4). Antibodies specific for these post-translational modifications are invaluable tools in the quest to understand normal and pathogenic molecular and cellular behavior.
General protein modification antibodies are designed to react with modified amino acid residues (e.g. phospho-threonine, phospho-tyrosine, acetyl-lysine, nitro-tyrosine) independently of the sequence in which they are embedded. This ability to recognize modified residues in a "context-independent" fashion gives these antibodies broad reactivities, presumably conferring upon them the ability to react with hundreds of distinct proteins. This broad pattern of reactivity makes these antibodies especially valuable in multiplex analyses and target discovery programs.
Protein kinases are among the most abundant eukaryotic regulatory proteins; over 500 separate kinase genes are encoded in mammalian genomes (5,6). In spite of the importance of kinases in eukaryotic biology, relatively few of their physiological targets are known. Phospho-Threonine Antibody (P-Thr-Polyclonal) #9381 and Phospho-Threonine (42H4) Monoclonal Antibody #9386 provide powerful tools for discovering targets of serine/threonine kinases, for monitoring and characterizing in vitro threonine phosphorylation reactions as well as for high throughput Ser/Thr kinase drug discovery.
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