CST, 84671T, SPAK (F7T1K) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying SPAK. Validated for WB,WB,IP,IHC,IF,IF. Available in 2 sizes. Highly specific and rigorously validated in-house, SPAK (F7T1K) Rabbit Monoclonal Antibody (CST #84671) is ready to ship. Product Usage Information Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:50 Immunohistochemistry (Paraffin): 1:400 - 1:1600 Immunofluorescence (Frozen): 1:400 - 1:800 Immunofluorescence (Immunocytochemistry): 1:400 - 1:1600 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry) Specificity / Sensitivity SPAK (F7T1K) Rabbit Monoclonal Antibody recognizes endogenous levels of total SPAK protein. Non-specific punctate cytoplasmic staining was observed in human kidney and staining of intercalated discs of human heart was observed by immunohistochemistry. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu429 of human SPAK protein. 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 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 Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 65 Source/Isotype: Rabbit IgG