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07-2273 Anti-phospho-SPAK Antibody (Ser373) / phospho-OSR1 Antibody (Ser325)

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 Blott

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Research Category: Signaling, Inflammation & Immunology Research Sub-Category: Kinases & Phosphatases, Oxidative Stress Gene Symbol: STK39, SPAK UniProt Number: Q9UEW8
07-2273
100 µg  
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Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
HWBRbAffinity PurifiedPolyclonal Antibody
Description
Catalogue Number07-2273
DescriptionAnti-phospho-SPAK Antibody (Ser373) / phospho-OSR1 Antibody (Ser325)
Alternate Names
  • 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 InformationSTE20/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.
Product Information
FormatAffinity Purified
Control
  • Hypotonic-stimulated HEK293 cells untreated and lambda phosphatase-treated.
PresentationPurified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationAnti-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
ImmunogenKLH-conjugated linear peptide corresponding to 15 amino acids surrounding phosphoserine 373 in human SPAK and phosphoserine 325 in human OSR1.
EpitopeSPAK phosphorlyated at Ser373 and OSR1 phosphorlyated at Ser325.
ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
HostRabbit
SpecificityThis 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 NoteHuman. Predicted to react with Mouse, Canine, Monkey, Porcine, Rat based on 100% sequence homology.
Antibody TypePolyclonal Antibody
Entrez Gene Number
Entrez Gene SummaryThis 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.
Gene Symbol
  • STK39
  • SPAK
Modifications
  • Phosphorylation
Purification MethodAffinity Purfied
UniProt Number
UniProt SummaryFUNCTION: May act as a mediator of stress-activated signals.

SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Note: Nucleus when caspase-cleaved.
Molecular WeightTarget 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).
Product Usage Statements
Quality AssuranceEvaluated 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.
Storage and Shipping Information
Storage ConditionsRecommended storage: +2°C to +8°C.
Packaging Information
Material Size100 µg
Global Trade Item Number
Catalogue Number GTIN
07-2273 04053252591211