Western blot analysis of extracts from various cell lines using Jak3 (5H2) Mouse mAb.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 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: 19
Jak3 (5H2) Mouse mAb detects endogenous levels of human Jak3 protein.
Monoclonal antibody was prepared from animals immunized with a purified recombinant fragment of human Jak3 expressed in E. Coli.
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.
Jak3 is primarily expressed in hematopoietic cells and is required for immune cell function and development (6-8). It binds to the common γ subunit (γc) and a shared receptor subunit also used by several cytokines including IL-2, IL-4, IL-7, IL-9, and IL-15 (9). IL-2 signaling and Stat5 activation is highly impaired by the loss of Jak3 (10,11). Jak3 is phosphorylated at multiple sites, including Tyr980 and 981 within its activation loop (12-14).
Explore pathways + proteins related to this product.