SPAK, Phosphorylated (Ser373), OSR1, Phosphorylated (Ser325) (STE20/SPS1-related Proline-alanine-rich Protein Kinase, Ste-20-related Kinase, DCHT, Serine/Threonine-protein Kinase 39, STK39/ Serine/Threonine-protein Kinase OSR1, Oxidative Stress-responsive

As members of the (sterile 20)-like kinase family, the oxidative-stress-responsive kinase 1 (OSR1) and the SPS1-related proline/alanine-rich kinase (SPAK) serve as key enzymes in a signaling cascade, which mediate the activity of Na+/K+/2Cl- cotransporters (NKCCs) in response to osmotic stress. Both kinases have a conserved carboxy-terminal (CCT) domain, which are 79% identical to each other at the sequence level, and are also highly conserved in orthologues of these enzymes in Caenorhabditis elegans, Drosophila, and Xenopus. The CCT domain reflects no similarity to sequences found in other proteins. This domain is known to recognize and interact with a unique peptide (Arg-Phe-Xaa-Val) motif, which is present in OSRT1- and SPAK-activating kinases (with-no-lysine-kinase 1 (WNK1 and WNK4) and also in STE20 kinase family substrates (NKCC1 and NKCC2). It is reported that phosphorylation of a Ser/Thr residue preceding the (Arg-Phe-Xaa-Val) motif results in steric clash, and subsequent dissociation from the CCT domain. Furthermore, patients suffering from Gordon syndrome carry mutated WNK1 and WNK4 genes.\n\nApplications:\nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 0.5ug/ml \nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nHypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

029614

Size

100ug

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Mouse

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 150mM sodium chloride, 0.05% sodium azide.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Villa, F., et al. (2007). EMBO Rep. 8(9):839-845. 2. Delpire, E. and Gagnon, K.B. (2008). 409(2): 321-331.

Additional Info

Recognizes human SPAK and OSR1 phosphorylated at Ser375 and Ser325 at ~60kD. Species Crossreactivity: mouse. Species Sequence Homology: rat and canine, 100%