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Anti-phospho-SPAK (Ser373)/phospho-OSR1 (Ser325), Cat. No. 07-2273, is a rabbit polyclonal antibody that detects phosphor SPAK (Ser373) and phospho-OSR1 (Ser325) and is tested for use in Western Blotting.
More>>Anti-phospho-SPAK (Ser373)/phospho-OSR1 (Ser325), Cat. No. 07-2273, is a rabbit polyclonal antibody that detects phosphor SPAK (Ser373) and phospho-OSR1 (Ser325) and is tested for use in Western Blotting. Less<<
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STE20/SPS1-related proline-alanine-rich protein kinase/Serine/threonine-protein kinase OSR1
EC:2.7.11.1
Ste-20-related kinase/Oxidative stress-responsive 1 protein
DCHT
Serine/threonine-protein kinase 39
Background Information
STE20/SPS1-related proline-alanine-rich protein kinase/Serine/threonine-protein kinase OSR1 (UniProt: Q9UEW8/O95747; also known as EC:2.7.11.1, Ste-20-related kinase/Oxidative stress-responsive 1 protein, DCHT, Serine/threonine-protein kinase 39) is encoded by the STK39/OXSR1 (also known as SPAK/OSR1) gene (Gene ID: 27347/9943) in human. SPAK and OSR1 are members of the (sterile 20)-like (STE20) kinase family. They serve as key enzymes in a signaling cascade, which mediate the activity of Na+/K+/2Cl- cotransporters (NKCCs) in response to osmotic stress. SPAK is predominantly expressed in brain and pancreas followed by heart, lung, kidney and skeletal muscle, liver, placenta, and testis. Its phosphorylated form is known to form a complex with WNK2. Its protein kinase domain is localized in amino acids 63-337. OSR1 is ubiquitously expressed, and its protein kinase domain resides in amino acids 17-291. Both kinases have a conserved carboxy-terminal (CCT) domain with 79% sequence homology. The CCT domain does not reflect similarity to sequences found in other proteins. This domain is known to recognize and interact with a unique peptide (Arg-Phe-Xaa-Val) motif that 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. Note: Please be aware that this antibody is called phospho-SPAK (Ser373) due to the historical notation of this post translational modification site over the last ~20 years of citations and publications. However, current UniProt sequence information for Q9UEW8-1 shows that this phosphorylation is actually located at Serine 371. It is currently unclear why this discrepancy exists. That said, we will continue to use the Serine 373 designation for this antibody, but researchers need to be aware of this inconsistency.
References
Product Information
Format
Affinity Purified
Control
Hypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.
Presentation
Purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-phospho-SPAK (Ser373)/phospho-OSR1 (Ser325), Cat. No. 07-2273, is a rabbit polyclonal antibody that detects phosphor SPAK (Ser373) and phospho-OSR1 (Ser325) and is tested for use in Western Blotting.
Key Applications
Western Blotting
Biological Information
Immunogen
KLH-conjugated linear peptide corresponding to 15 amino acids surrounding phosphoserine 373 in human SPAK and phosphoserine 325 in human OSR1.
Epitope
SPAK phosphorlyated at Ser373 and OSR1 phosphorlyated at Ser325.
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Rabbit
Specificity
This rabbit polyclonal antibody detects phospho-SPAK (Ser373)/phospho-OSR1 (Ser325). Note: Please be aware that this antibody is called phospho-SPAK (Ser373) due to the historical notation of this post translational modification site over the last ~20 years of citations and publications. However, current UniProt sequence information for Q9UEW8-1 shows that this phosphorylation is actually located at Serine 371. It is currently unclear why this discrepancy exists. That said, we will continue to use the Serine 373 designation for this antibody, but researchers need to be aware of this inconsistency.
Species Reactivity
Human
Species Reactivity Note
Human. Predicted to react with Mouse, Canine, Monkey, Porcine, Rat based on 100% sequence homology.
This gene encodes a serine/threonine kinase that is thought to function in the cellular stress response pathway. The kinase is activated in response to hypotonic stress, leading to phosphorylation of several cation-chloride-coupled cotransporters. The catalytically active kinase specifically activates the p38 MAP kinase pathway, and its interaction with p38 decreases upon cellular stress, suggesting that this kinase may serve as an intermediate in the response to cellular stress.
FUNCTION: May act as a mediator of stress-activated signals.
SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Note: Nucleus when caspase-cleaved.
Molecular Weight
Target molecular weight ~59 and 60 kDa observed; 59.47 and 58.02 kDa calculated, respectively for SPAK and OSR1. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in lysate from HEK293 cells stimulated with hypotonic buffer.
Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected phospho-SPAK (Ser373)/ phospho-OSR1 (Ser325) in HEK293 cells stimulated with a hypotonic buffer. However, this antibody does not recognize SPAK/OSR1 protein treated with protein phosphatase.
Usage Statement
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.
