Anti-phospho-SPAK Antibody (Ser373) / phospho-OSR1 Antibody (Ser325)

General description

As members of the (sterile 20)-like (STE20) 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.

Specificity

This antibody recognizes SPAK phosphorlyated at Ser373 and OSR1 phosphorlyated at Ser325.

Immunogen

Epitope: SPAK phosphorlyated at Ser373 and OSR1 phosphorlyated at Ser325.

KLH-conjugated linear peptide corresponding to human SPAK and OSR1 phosphorylated at Ser375 and Ser325, respectively.

Application

Research Category
Signaling

Inflammation & Immunology

Research Sub Category
Kinases & Phosphatases

Oxidative Stress

This phospho-SPAK antibody is validated for use in WB for the detection of the phospho-SPAK protein.

Quality

Evaluated by Western Blot in hypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.

Western Blot Anlaysis: 0.5 µg/mL of this antibody detected SPAK / OSR1 in 10 µg hypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.

Target description

~60 kDa observed

Physical form

Affinity purified

Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Analysis Note

Control
Hypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.