SPAK Antibody

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WB
IF

Inquiry Info. # 2281

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Product Specifications

REACTIVITY	H M R Mk
SENSITIVITY	Endogenous
MW (kDa)	65
SOURCE	Rabbit

Application Key:

WB-Western Blotting
IF-Immunofluorescence

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat
Mk-Monkey

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunofluorescence (Frozen)	1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Protocol

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.

Species Reactivity:

Human, Mouse, Rat, Monkey

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.

Background

SPAK (STE20/SPS1-related Pro/Ala-rich kinase) and OSR1 (oxidative stress responsive 1) are members of the GCK family of 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 knockdown of SPAK dramatically reduces TCR/CD28-induced AP-1 activation, suggesting SPAK is involved in T cell signaling as well (6).

Alternate Names

DCHT; DKFZp686K05124; PASK; proline-alanine-rich STE20-related kinase; serine threonine kinase 39 (STE20/SPS1 homolog, yeast); serine/threonine kinase 39; Serine/threonine-protein kinase 39; small intestine SPAK-like kinase; SPAK; Ste-20-related kinase; Ste20-like protein kinase; STE20/SPS1 homolog; STE20/SPS1-related proline-alanine-rich protein kinase; STK39; STLK3

Database Links

UniProt ID: Q9UEW8

Entrez-Gene Id: 27347

Limited Uses

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SPAK Antibody #2281