Phospho-WNK1 (T60) and Total WNK1 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-WNK1 (T60) and Total WNK1 in Human in Cell Lysate samples.
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Phospho-WNK1 (T60) and Total WNK1 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-WNK1 (T60) and Total WNK1 in Human in Cell Lysate samples.
Phospho-WNK1 (T60) and Total WNK1 ELISA Kit (ab279969) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated WNK1 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-WNK1 and total WNK1. An anti-pan WNK1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and WNK1 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho WNK1 (T60) antibody is added to detect phosphorylated WNK1. In the remaining wells, rabbit anti-pan-WNK1 antibody is used to detect pan WNK1. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of WNK1 (T60) or pan WNK1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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WNK1 also known as With No Lysine 1 is a protein kinase with a molecular mass of approximately 251 kDa. Mechanically WNK1 functions as a serine/threonine kinase influencing the phosphorylation status of various substrates involved in ionic balance and cell volume regulation. It plays an important role in controlling ion transport channels. The protein is expressed in multiple tissues with high levels in the kidney and lower levels in the brain heart and other regions of the body. Its activity is especially important in the distal nephrons of the kidney.
WNK1 affects sodium and potassium ion transport which is essential for maintaining homeostasis. It is a part of a larger signaling complex where it modulates the activity of SPAK and OSR1 kinases which directly affect ion transporters like NKCCs and NCC. By regulating these kinases WNK1 contributes to maintaining the balance of electrolytes influencing blood pressure and general cellular osmoregulation.
WNK1 significantly impacts the WNK-SPAK/OSR1 signaling pathway which has direct roles in sodium and potassium ion transport. The pathway associates closely with proteins like SPAK and OSR1 which WNK1 activates affecting the function of ion transporters and channels. This signaling mechanism plays an integral role in renal salt reabsorption and is important for controlling systemic blood pressure.
Mutations and dysregulation of WNK1 link to hereditary hypertension and pseudohypoaldosteronism type II (PHAII) also known as Gordon's syndrome. These conditions involve elevated blood pressure and electrolyte imbalance due to altered transport activity. A change in WNK1 function can disrupt normal interaction with other proteins in its pathway such as NCC leading to these disorders. Understanding WNK1's role offers potential therapeutic targets for managing conditions related to electrolyte imbalance and blood pressure regulation.
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Terms & Conditions.
Positive Control.
T47D cells were treated with IGF-1 at 37°C for 30 min.
Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
HeLa cells treated with/without IGF-1.
HeLa cells were untreated or treated with IGF-1 (50 ng/ml) for 30 min.
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