Product Name:
WNK1-pT60
Product Number:
ab-pk856
Target Full Name: Serine/threonine-protein kinase WNK1
Target Alias: HSAN2; HSN2; KDP; KIAA0344; MGC163339; MGC163341; p65; PRKWNK1; Protein kinase with no lysine 1; Protein kinase, lysine-deficient 1; PSK; PHA2C; CCDS8506.1; ENSG00000060237
Product Type Specific: WNK1 protein kinase phosphosite-specific antibody
Antibody Code: PK856
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T60
Protein UniProt: Q9H4A3
Protein SigNET: WNK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human WNK1 sequence peptide Cat. No.: PE-04ALJ99
Antibody Immunogen Sequence: RRH(pT)MDK(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues R57 to K63; in the N-terminal portion of the protein kinase. T60 phosphorylation stimulates phosphotransferase activity. This is the major site of in vivo phosphorylation of WNK1 based on ≥211 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus. WNK1 is known to be phosphorylated at this site in vitro by Akt1 (PKBa), Akt3 (PKBg), MSK1 (RPS6KA5), and SGK1.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Dog | Cow | Rat | Mouse | Chicken.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 190, 250-400 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: Almost no significant cross-reactivities detected in A431 and Jurkat cells, except for a 75 kDa cross-reactive protein that is also detected in sea star oocytes. Sea star oocytes also feature 140 and 200 kDa strongly cross-reactive proteins.
Related Product 1: WNK1 (57-63) pT60 antibody blocking peptide (Cat. No.: PE-04ALJ99)
Related Product 2: WNK1-1 pan-specific antibody (Cat. No.: AB-NK252-1)
Related Product 3: WNK1-2 pan-specific antibody (Cat. No.: AB-NK252-2)
Related Product 4: WNK1-3 pan-specific antibody (Cat. No.: AB-NK252-3)
Related Product 5: WNK1-pS382 phosphosite-specific antibody (Cat. No.: AB-PK855)
Related Product 6: WNK1-pT2245 phosphosite-specific antibody (Cat. No.: AB-PK857)
Scientific Background: WNK1 (With No Lysine 1); PRKWNK1) is a 230 kDa protein-serine/threonine kinase of the Other group and WNK family. It functions as a key osmotic sensor and regulator of blood pressure, ion transport (sodium, potassium, chloride), and pain sensation. It functions primarily in the distal nephron to regulate ion channels. It is involved in electrolyte homeostasis, cell survival, cell signalling, and cell proliferation. It is activated by hypertonicity. The activation requires autophosphorylation of Ser-382. Phosphorylation of Ser-378 also promotes increased phosphotransferase activity. WNK1 can activate sodium-coupled chloride cotransporters, and inhibit potassium-coupled chloride cotransporters. WNK1 activates NKCC1, NKCC2, and NCC, while inhibiting the ROMK1 potassium channel to regulate blood pressure and electrolyte homeostasis. Other proteins activated by WNK1 include SCNN1A, SCNN1B, SCNN1D and SGK1 while another inhibited protein is WNK4 (via phosphorylation of WNK4 or an associated complex of a WNK1 autoinhibitory domain bound to WNK4). WNK4 inhibition could modulate, via phosphorylation, a thiazide-sensitive NaCl cotransporter transporter (specifically SLC12A3). WNK1 has also been proposed to modulate NEDD4L phosphorylation, and have an effect on actin cytoskeleton rearrangement. As an "atypical" kinase, WNK1 lacks a standard catalytic lysine in subdomain II, utilizing a different mechanism (a kinase domain in the N-terminus) to bind ATP, and acts as a kinase for SPAK and OXSR1 kinases. It exists in multiple forms, including the ubiquitous long-WNK1 (L-WNK1) and the kidney-specific kinase-deficient isoform, KS-WNK1, which together balance potassium and sodium transport. WNK1 is found in the kidneys, heart, skeletal muscle, and the nervous system (DRG neurons). Its overactivity resulting in pseudohypoaldosteronism type II, a disease characterized by salt-sensitive hypertension and hyperkalemia. Insertional mutagenesis studies in mice support a role for this protein kinase in mouse cancer oncogenesis. This description may include information annotated by UniProt and/or Google AI.