Western blot analysis of extracts from HT-29 cells treated with IL-4 (100 ng/ml) for the indicated times, using Phospho-Jak1 (Tyr1022/1023) Antibody (upper) or Jak1 Antibody (lower).
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-Jak1 (Tyr1034/1035) Antibody detects endogenous levels of Jak1 only when phosphorylated at tyrosines 1034/1035. Human Jak1 residues Tyr1034 and Tyr1035 historically have been referenced as Tyr1022 and Tyr1023. This antibody may cross-react with phospho-Jak2.
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1034/1035 of human Jak1. Antibodies are purified by protein A and peptide affinity chromatography.
Members of the Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) are activated by ligands binding to a number of associated cytokine receptors (1). Upon cytokine receptor activation, Jak proteins become autophosphorylated and phosphorylate their associated receptors to provide multiple binding sites for signaling proteins. These associated signaling proteins, such as Stats (2), Shc (3), insulin receptor substrates (4), and focal adhesion kinase (FAK) (5), typically contain SH2 or other phospho-tyrosine-binding domains.
The tyrosine residues 1022/1023 of Jak1 in the putative activation loop are the homologous tyrosine residues 1054/1055 in Tyk2, which are important in the regulation of Tyk2 kinase activity (6).
(This product is sold under license from Chemicon, Inc., U.S. Patent No. 5,658,791.)
Explore pathways + proteins related to this product.
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