SPAK (STE20/SPS1-related Pro/Ala-rich kinase) and OSR1 (oxidative stress responsive 1) are members of the GCK family serine/threonine kinases.
Overexpression and in vitro studies demonstrate that SPAK is able to activate p38 MAP kinase indicating a possible role for SPAK in the stress response.
Yeast two-hybrid screening revealed that SPAK and OSR1 bind to Na-K-2Cl cotransporters NKCC1 and NKCC2 and K-Cl cotransporter KCC3 (2).
WNK1 and WNK4 phosphorylate SPAK at Thr243/247 and Ser380.
Similarly, WNK1 and WNK4 phosphorylate OSR1 at Thr185 and Ser315.
Phosphorylation at these sites stimulates SPAK and OSR1 activity, leading to NKCC1 phosphorylation and enhanced NKCC1 activity.
SPAK is also phosphorylated at Ser311 by PKC theta in response to T cell activation.
Substitution of Ser311 with Ala or specific siRNA knock-down of SPAK dramatically reduces TCR/CD28-induced AP-1 activation, suggesting SPAK is involved in T cell signaling as well.
Applications:Suitable for use in Western Blot, Flow Cytometry and ImmunofluorescenceI/mmunocytochemistry.
Other applications not tested.
Recommended Dilutions:Western Blot: 1:1000.
Incubate membrane with diluted antibody in TBS, 5% BSA, 0.1% Tween-20 at 4°C with gentle shaking, overnight
Flow Cytometry: 1:100Immunofluorescence (IC): 1:25Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A and peptide affinity chromatography.
Form：Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity：Recognizes endogenous levels of total human OSR1 at ~60kD. Does not crossreact with SPAK or other members of the GCK family. Species Crossreactivity: Bovine and monkey
Isotype：IgG
Calc Applications Abbrev：FC IC IF WB
Calc Crossreactivity：Bo Hu Mk
Immunogen：Synthetic peptide corresponding to residues at the N-terminus of human OSR1.
