WNK1 (Phospho-Thr58) ELISA Kit

Name: WNK1 (Phospho-Thr58) ELISA Kit
SKU: DEIA-XYA1831
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

cultured cells

Species Reactivity

Human; Mouse; Rat

Intended Use

The WNK1 (Phospho-Thr58) Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can monitor WNK1 protein phosphorylation and expression profile in cells. The kit can be used for measuring the relative amounts of phosphorylated WNK1 in cultured cells as well as screening for the effects that various treatments, inhibitors (ie. siRNA or chemicals), or activators have on WNK1 phosphorylation.

Contents of Kit

1. 96-Well Cell Culture Clear-Bottom Microplate: 2 Plates
2. 10x TBS: 24 mL (10x), Clear
3. Quenching Buffer: 24 mL (1x), Clear
4. Blocking Buffer: 50 mL (1x), Clear
5. 10x Wash Buffer: 50 mL (10x), Clear
6. 100x Anti-WNK1 (Phospho-Thr58) Antibody (Rabbit Polyclonal): 60 μL (100x), Red
7. 100x Anti-WNK1 Antibody (Rabbit Polyclonal): 60 μL (100x), Purple
8. 100x Anti-GAPDH Antibody (Mouse Monoclonal): 60 μL (100x), Green
9. HRP-Conjugated Anti-Rabbit IgG Antibody: 12 mL (1x), Glass
10. HRP-Conjugated Anti-Mouse IgG Antibody: 12 mL (1x), Glass
11. Primary Antibody Diluent: 12 mL (1x), Clear
12. Ready-to-Use Substrate: 12 mL (1x), Brown
13. Stop Solution: 12 mL (1x), Clear
14. Crystal Violet Solution: 12 mL (1x), Glass
15. SDS Solution: 24 mL (1x), Clear
16. Adhesive Plate Seals: 4 Seals

Storage

4°C/6 Months

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Association of with-no-lysine kinase 1 and Serine/Threonine kinase 39 gene polymorphisms and haplotypes with essential hypertension in Tibetans

ENVIRONMENTAL AND MOLECULAR MUTAGENESIS

Authors: Shi, Rufeng; Li, Jiangbo; He, Jiyun; Meng, Qingtao; Qian, Zhiping; Shi, Di; Liu, Qi; Cai, Yali; Li, Xinran; Chen, Xiaoping

Abstract

Tibetans have a higher essential hypertension prevalence compared with other ethnics in China. The reason might be due to their unique environmental influence, as well as genetic factor. However, limited studies focus on Tibetan genetics and its association with hypertension. The aim of this study was to investigate the association between With-No-Lysine (K) Kinase 1 (WNK1), Serine/Threonine kinase 39(STK39) genes variants and hypertension in the Tibetan population. 204 Tibetan hypertensive patients and 305 normotensive controls were recruited in an epidemiological survey conducted at 2 sites in the Ganzi Tibetan autonomous region. Patients were genotyped for nineteen WNK1 candidate tag single nucleotide polymorphisms (SNPs) and three STK39 SNPs, and haplotype analysis was performed. Results showed that the allele A in rs1468326 was overrepresented in hypertensive patients versus control (53.4% vs 42.9%, P<0.05). The multivariable-adjusted odds ratio (OR) for hypertension among CA+AA genotypes carriers was 1.60 (95% CI: 1.02-2.62, P<0.05), and they also had a higher systolic blood pressure (136.5 +/- 28.6 vs 131.7 +/- 24.8 mmHg, P<0.05). However, the TT genotype ratio in rs6749447 was lower in hypertensives (5.4% vs 10.8%, P<0.05), and the hypertension risk for the TT genotype carriers in rs6749447 decreased after adjustment (OR 0.49, 95% CI 0.19-0.95, P<0.05). Subjects with haplotype AGACAGGAATCGT showed 1.57 times higher risk of hypertension (95% CI 1.02-2.41, P<0.05). In conclusion, SNP rs1468326 of WNK1, rs6749447 of STK39, and WNK1 haplotype AGACAGGAATCGT were associated with hypertension in Tibetan individuals. Environ. Mol. Mutagen. 59:151-160, 2018. (c) 2017 Wiley Periodicals, Inc.

Intersectin links WNK kinases to endocytosis of ROMK1

JOURNAL OF CLINICAL INVESTIGATION

Authors: He, Guocheng; Wang, Hao-Ran; Huang, Shao-Kuei; Huang, Chou-Long

Abstract

With-no-lysine (WNK) kinases are a novel family of protein kinases characterized by an atypical placement of the catalytic lysine. Mutations of 2 family members, WNK1 and WNK4, cause pseudohypoaldosteronism type 2 (PHA2), an autosomal-dominant disease characterized by hypertension and hyperkalemia. WNK1 and WNK4 stimulate clathrin-dependent endocytosis of renal outer medullar potassium 1 (ROMK1), and PHA2-causing mutations of WNK4 increase the endocytosis. How WNKs stimulate endocytosis of ROMK1 and how mutations of WNK4 increase the endocytosis are unknown. Intersectin (ITSN) is a multimodular endocytic scaffold protein. Here we show that WNK1 and WNK4 interacted with ITSN and that the interactions were crucial for stimulation of endocytosis of ROMK1 by WNKs. The stimulation of endocytosis of ROMK1 by WNK1 and WNK4 required specific proline-rich motifs of WNKs, but did not require their kinase activity. WNK4 interacted with ROMK1 as well as with ITSN. Disease-causing WNK4 mutations enhanced interactions of WNK4 with ITSN and ROMK1, leading to increased endocytosis of ROMK1. These results provide a molecular mechanism for stimulation of endocytosis of ROMK1 by WNK kinases.