Phospho-WNK1 (Thr60) Antibody - #AF3483

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Product: Phospho-WNK1 (Thr60) Antibody
Catalog: AF3483
Description: Rabbit polyclonal antibody to Phospho-WNK1 (Thr60)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 230kDa; 251kD(Calculated).
Uniprot: Q9H4A3
RRID: AB_2834921

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 100ul $280 In stock
 200ul $350 In stock

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
Phospho-WNK1 (Thr60) Antibody detects endogenous levels of WNK1 only when phosphorylated at Threonine 60.
RRID:
AB_2834921
Cite Format: Affinity Biosciences Cat# AF3483, RRID:AB_2834921.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Erythrocyte 65 kDa protein; HSAN2; HSN2; hWNK1; KDP; KIAA0344; Kinase deficient protein; MGC163339; MGC163341; p65; PPP1R167; PRKWNK1; Prostate derived sterile 20 like kinase; Protein kinase lysine deficient 1; Protein kinase lysine-deficient 1; Protein kinase with no lysine 1; Protein phosphatase 1, regulatory subunit 167; PSK; Serine/threonine protein kinase WNK1 1; Serine/threonine protein kinase WNK1 2; Serine/threonine protein kinase WNK1; Serine/threonine-protein kinase WNK1; With no K; WNK lysine deficient protein kinase 1; WNK lysine deficient protein kinase 1 isoform; WNK1; WNK1_HUMAN;

Immunogens

Immunogen:
Uniprot:

Q9H4A3(WNK1_HUMAN) >>Visit HPA database.

Gene(ID):
WNK1(65125)
Expression:
Q9H4A3 WNK1_HUMAN:

Widely expressed, with highest levels observed in the testis, heart, kidney and skeletal muscle. Isoform 3 is kidney-specific and specifically expressed in the distal convoluted tubule (DCT) and connecting tubule (CNT) of the nephron.

Description:
WNK1 a serine-threonine protein kinase that controls sodium and chloride ion transport. May regulate the activity of the thiazide-sensitive Na-Cl cotransporter SLC12A3 by phosphorylation. May also play a role in actin cytoskeletal reorganization.
Sequence:
MSGGAAEKQSSTPGSLFLSPPAPAPKNGSSSDSSVGEKLGAAAADAVTGRTEEYRRRRHTMDKDSRGAAATTTTTEHRFFRRSVICDSNATALELPGLPLSLPQPSIPAAVPQSAPPEPHREETVTATATSQVAQQPPAAAAPGEQAVAGPAPSTVPSSTSKDRPVSQPSLVGSKEEPPPARSGSGGGSAKEPQEERSQQQDDIEELETKAVGMSNDGRFLKFDIEIGRGSFKTVYKGLDTETTVEVAWCELQDRKLTKSERQRFKEEAEMLKGLQHPNIVRFYDSWESTVKGKKCIVLVTELMTSGTLKTYLKRFKVMKIKVLRSWCRQILKGLQFLHTRTPPIIHRDLKCDNIFITGPTGSVKIGDLGLATLKRASFAKSVIGTPEFMAPEMYEEKYDESVDVYAFGMCMLEMATSEYPYSECQNAAQIYRRVTSGVKPASFDKVAIPEVKEIIEGCIRQNKDERYSIKDLLNHAFFQEETGVRVELAEEDDGEKIAIKLWLRIEDIKKLKGKYKDNEAIEFSFDLERDVPEDVAQEMVESGYVCEGDHKTMAKAIKDRVSLIKRKREQRQLVREEQEKKKQEESSLKQQVEQSSASQTGIKQLPSASTGIPTASTTSASVSTQVEPEEPEADQHQQLQYQQPSISVLSDGTVDSGQGSSVFTESRVSSQQTVSYGSQHEQAHSTGTVPGHIPSTVQAQSQPHGVYPPSSVAQGQSQGQPSSSSLTGVSSSQPIQHPQQQQGIQQTAPPQQTVQYSLSQTSTSSEATTAQPVSQPQAPQVLPQVSAGKQLPVSQPVPTIQGEPQIPVATQPSVVPVHSGAHFLPVGQPLPTPLLPQYPVSQIPISTPHVSTAQTGFSSLPITMAAGITQPLLTLASSATTAAIPGVSTVVPSQLPTLLQPVTQLPSQVHPQLLQPAVQSMGIPANLGQAAEVPLSSGDVLYQGFPPRLPPQYPGDSNIAPSSNVASVCIHSTVLSPPMPTEVLATPGYFPTVVQPYVESNLLVPMGGVGGQVQVSQPGGSLAQAPTTSSQQAVLESTQGVSQVAPAEPVAVAQTQATQPTTLASSVDSAHSDVASGMSDGNENVPSSSGRHEGRTTKRHYRKSVRSRSRHEKTSRPKLRILNVSNKGDRVVECQLETHNRKMVTFKFDLDGDNPEEIATIMVNNDFILAIERESFVDQVREIIEKADEMLSEDVSVEPEGDQGLESLQGKDDYGFSGSQKLEGEFKQPIPASSMPQQIGIPTSSLTQVVHSAGRRFIVSPVPESRLRESKVFPSEITDTVAASTAQSPGMNLSHSASSLSLQQAFSELRRAQMTEGPNTAPPNFSHTGPTFPVVPPFLSSIAGVPTTAAATAPVPATSSPPNDISTSVIQSEVTVPTEEGIAGVATSTGVVTSGGLPIPPVSESPVLSSVVSSITIPAVVSISTTSPSLQVPTSTSEIVVSSTALYPSVTVSATSASAGGSTATPGPKPPAVVSQQAAGSTTVGATLTSVSTTTSFPSTASQLCIQLSSSTSTPTLAETVVVSAHSLDKTSHSSTTGLAFSLSAPSSSSSPGAGVSSYISQPGGLHPLVIPSVIASTPILPQAAGPTSTPLLPQVPSIPPLVQPVANVPAVQQTLIHSQPQPALLPNQPHTHCPEVDSDTQPKAPGIDDIKTLEEKLRSLFSEHSSSGAQHASVSLETSLVIESTVTPGIPTTAVAPSKLLTSTTSTCLPPTNLPLGTVALPVTPVVTPGQVSTPVSTTTSGVKPGTAPSKPPLTKAPVLPVGTELPAGTLPSEQLPPFPGPSLTQSQQPLEDLDAQLRRTLSPEMITVTSAVGPVSMAAPTAITEAGTQPQKGVSQVKEGPVLATSSGAGVFKMGRFQVSVAADGAQKEGKNKSEDAKSVHFESSTSESSVLSSSSPESTLVKPEPNGITIPGISSDVPESAHKTTASEAKSDTGQPTKVGRFQVTTTANKVGRFSVSKTEDKITDTKKEGPVASPPFMDLEQAVLPAVIPKKEKPELSEPSHLNGPSSDPEAAFLSRDVDDGSGSPHSPHQLSSKSLPSQNLSQSLSNSFNSSYMSSDNESDIEDEDLKLELRRLRDKHLKEIQDLQSRQKHEIESLYTKLGKVPPAVIIPPAAPLSGRRRRPTKSKGSKSSRSSSLGNKSPQLSGNLSGQSAASVLHPQQTLHPPGNIPESGQNQLLQPLKPSPSSDNLYSAFTSDGAISVPSLSAPGQGTSSTNTVGATVNSQAAQAQPPAMTSSRKGTFTDDLHKLVDNWARDAMNLSGRRGSKGHMNYEGPGMARKFSAPGQLCISMTSNLGGSAPISAASATSLGHFTKSMCPPQQYGFPATPFGAQWSGTGGPAPQPLGQFQPVGTASLQNFNISNLQKSISNPPGSNLRTT

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Chicken
100
Rabbit
100
Horse
0
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q9H4A3 As Substrate

Site PTM Type Enzyme
S2 Phosphorylation
S11 Phosphorylation
T12 Phosphorylation
S15 Phosphorylation
S19 Phosphorylation
S33 Phosphorylation
T60 Phosphorylation O00141 (SGK1) , P31749 (AKT1) , O75582 (RPS6KA5)
R66 Methylation
T74 Phosphorylation
S167 Phosphorylation
S170 Phosphorylation
S174 Phosphorylation
S183 Phosphorylation
S185 Phosphorylation
S189 Phosphorylation
S198 Phosphorylation
S231 Phosphorylation
S306 Phosphorylation
T308 Phosphorylation
T373 Phosphorylation
K375 Ubiquitination
S378 Phosphorylation Q9H4A3 (WNK1)
S382 Phosphorylation Q9BYP7 (WNK3) , Q9Y3S1 (WNK2) , Q9H4A3 (WNK1)
S599 Phosphorylation
S679 Phosphorylation
T1098 Phosphorylation
Y1102 Phosphorylation
K1104 Acetylation
S1105 Phosphorylation
S1108 Phosphorylation
S1110 Phosphorylation
T1115 Phosphorylation
S1116 Phosphorylation
K1119 Acetylation
K1128 Acetylation
Y1215 Phosphorylation
S1220 Phosphorylation
S1253 Phosphorylation
S1261 Phosphorylation
S1285 Phosphorylation
S1295 Phosphorylation
S1297 Phosphorylation
S1300 Phosphorylation
K1653 Ubiquitination
K1658 Ubiquitination
T1810 Phosphorylation
T1848 Phosphorylation
S1849 Phosphorylation
S1887 Phosphorylation
S1935 Phosphorylation
S1959 Phosphorylation
S1961 Phosphorylation
T1963 Phosphorylation
T1968 Phosphorylation
T1970 Phosphorylation
S1978 Phosphorylation
S2002 Phosphorylation
S2005 Phosphorylation
S2011 Phosphorylation
S2012 Phosphorylation
S2027 Phosphorylation
S2029 Phosphorylation
S2032 Phosphorylation P42345 (MTOR)
S2037 Phosphorylation
S2038 Phosphorylation
S2065 Phosphorylation
Y2102 Phosphorylation
S2121 Phosphorylation
T2128 Phosphorylation
S2145 Phosphorylation
K2243 Ubiquitination
T2245 Phosphorylation
T2247 Phosphorylation
S2265 Phosphorylation
S2270 Phosphorylation
Y2276 Phosphorylation
S2286 Phosphorylation
S2294 Phosphorylation
S2370 Phosphorylation
S2372 Phosphorylation
R2380 Methylation

PTMs - Q9H4A3 As Enzyme

Substrate Site Source
O95747 (OXSR1) T185 Uniprot
O95747 (OXSR1) S325 Uniprot
Q15796 (SMAD2) S110 Uniprot
Q15796 (SMAD2) S260 Uniprot
Q15796 (SMAD2) S465 Uniprot
Q8N9I0 (SYT2) T199 Uniprot
Q8N9I0 (SYT2) T383 Uniprot
Q9H4A3 (WNK1) S378 Uniprot
Q9H4A3 (WNK1) S382 Uniprot
Q9UEW8 (STK39) T231 Uniprot
Q9UEW8 (STK39) S371 Uniprot
Q9UHW9 (SLC12A6) T991 Uniprot
Q9UHW9 (SLC12A6) T1048 Uniprot

Research Backgrounds

Function:

Serine/threonine kinase which plays an important role in the regulation of electrolyte homeostasis, cell signaling, survival, and proliferation. Acts as an activator and inhibitor of sodium-coupled chloride cotransporters and potassium-coupled chloride cotransporters respectively. Activates SCNN1A, SCNN1B, SCNN1D and SGK1. Controls sodium and chloride ion transport by inhibiting the activity of WNK4, by either phosphorylating the kinase or via an interaction between WNK4 and the autoinhibitory domain of WNK1. WNK4 regulates the activity of the thiazide-sensitive Na-Cl cotransporter, SLC12A3, by phosphorylation. WNK1 may also play a role in actin cytoskeletal reorganization. Phosphorylates NEDD4L. Acts as a scaffold to inhibit SLC4A4, SLC26A6 as well as CFTR activities and surface expression, recruits STK39 which mediates the inhibition (By similarity).

Dominant-negative regulator of the longer isoform 1. Does not have kinase activity, does not directly inhibit WNK4 and has no direct effect on sodium and chloride ion transport. Downregulates sodium-chloride cotransporter activity indirectly by inhibiting isoform 1, it associates with isoform 1 and attenuates its kinase activity. In kidney, may play an important role regulating sodium and potassium balance.

PTMs:

Autophosphorylation at Ser-382 is inhibited by intracellular calcium.

May be O-glycosylated.

Ubiquitinated in vitro by the BCR(KLHL3) complex and in vivo by a BCR(KLHL2) complex, leading to proteasomal degradation.

Subcellular Location:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed, with highest levels observed in the testis, heart, kidney and skeletal muscle. Isoform 3 is kidney-specific and specifically expressed in the distal convoluted tubule (DCT) and connecting tubule (CNT) of the nephron.

Subunit Structure:

Interacts with SYT2 (By similarity). Interacts with WNK3 and WNK4 (By similarity). Interacts with KLHL3. Isoform 3: Interacts with isoform 1 (By similarity).

Family&Domains:

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. WNK subfamily.

References

1). Regulatory role of PI3K/Akt/WNK1 signal pathway in mouse model of bone cancer pain. Scientific reports, 2023 (PubMed: 37652923) [IF=3.8]

Application: WB    Species: Mouse    Sample:

Figure 6 (A)、(C)、(E) Western blotting to detect the expression of PI3K, p-PI3K, Akt, p-Akt, WNK1 and p-WNK1 protein. (B)、(D)、(F) The optical density of protein bands detected by Western blotting was statistically quantified by IPP 6.0 software; (B) Protein expression of PI3K and p-PI3K; (D) Protein expression of Akt and p-Akt; (F) Protein expression of WNK1 and p-WNK1. N: Normal group, BCP: Bone cancer pain group, Akt-i: Akt inhibitor group. n = 3; *P 

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