Product Pathways - Cytoskeletal Signaling
SPAK Antibody #2281
|W IF-F||H M R Mk||Endogenous||65||Rabbit|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
SPAK Antibody detects endogenous levels of total SPAK protein. This antibody does not cross-react with OSR1 or other members of the GCK family.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser436 of human SPAK. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from 293, C2C12 and NBT-II cells, using SPAK Antibody.
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 (1). 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 (3-5). Similarly, WNK1 and WNK4 phosphorylate OSR1 at Thr185 and Ser315 (3, 4). Phosphorylation at these sites stimulates SPAK and OSR1 activity, leading to NKCC1 phosphorylation and enhanced NKCC1 activity (3-5). SPAK is also phosphorylated at Ser311 by PKCθ 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 (6).
- Johnston, A.M. et al. (2000) Oncogene 19, 4290-4297.
- Piechotta, K. et al. (2002) J. Biol. Chem. 277, 50812-50819.
- Vitari, A.C. et al. (2005) Biochem. J. 391, 17-24.
- Moriguchi, T. et al. (2005) J. Biol. Chem. 280, 42685-42693.
- Gagnon, K.B. et al. (2006) Mol. Cell. Biol. 26, 689-698.
- Li, Y. et al. (2004) EMBO J. 23, 1112-1122.
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For Research Use Only. Not For Use In Diagnostic Procedures.