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Anti ncc

Manufactured by Merck Group
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Sourced in Germany, United Kingdom

Anti-NCC is a laboratory equipment product designed for the detection and quantification of the NCC (Sodium-Coupled Cotransporter) protein. It is a specialized tool used in research and clinical settings to study the expression and activity of NCC, a key regulator of sodium and water balance in the body.

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Lab products found in correlation

Anti β actin, by Merck Group (2 mentions) Anti wnk4, by Novus Biologicals (2 mentions) Anti aqp2, by Santa Cruz Biotechnology (1 mentions) Anti cav 1, by Santa Cruz Biotechnology (1 mentions) Anti enos, by Santa Cruz Biotechnology (1 mentions) Anti nkcc1, by Developmental Studies Hybridoma Bank (1 mentions) Anti na k atpase, by Merck Group (1 mentions) Anti aqp1, by Alpha Diagnostic (1 mentions) Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions) Imagequant 5, by GE Healthcare (1 mentions) Anti α enac, by Novus Biologicals (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Anti phosphoserine, by Alpha Diagnostic (1 mentions) Protein a or g agarose sepharose beads, by Santa Cruz Biotechnology (1 mentions) Plasmamembrane isolation kit, by Invent Biotechnologies (1 mentions) Anti ubiquitin, by Cell Signaling Technology (1 mentions) Anti wnk1, by Cell Signaling Technology (1 mentions) Anti wnk4, by Merck Group (1 mentions) Anti phospho spak osr1, by Merck Group (1 mentions) Anti calcineurin a, by Cell Signaling Technology (1 mentions)

4 protocols using anti ncc

1

Immunostaining of Aquaporin Proteins

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The following primary antibodies were applied for immunofluorescence, immunohistochemistry, or immunoblotting: anti-AQP1 (Alpha diagnostic international, San Antonio, Texas, USA), anti-AQP2 (Santa Cruz Biotechnology, Heidelberg, Germany), anti-phospho-aquaporin 2 (pS256)55 , anti-β-actin (Sigma-Aldrich, St. Louis, USA), anti-Cav1 (Santa Cruz Biotechnology, Heidelberg, Germany), anti-NKCC1 (T4; Developmental Studies Hybridoma Bank, University of Iowa, USA), anti-vasopressin V2 receptor56 (link), anti-eNOS (Santa Cruz Biotechnology), anti-Na+/K+-ATPase (Millipore, Darmstadt, Germany), anti-NCC, anti-NKCC2, anti-phospho-NKCC2 (pT96/pT101), and anti-phospho-NCC (pS71)57 (link).
Willière Y., Borschewski A., Patzak A., Nikitina T., Dittmayer C., Daigeler A.L., Schuelke M., Bachmann S, & Mutig K. (2018). Caveolin 1 Promotes Renal Water and Salt Reabsorption. Scientific Reports, 8, 545.
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2

Immunoblot Analysis of Kidney Proteins

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Kidney cortex homogenates or total cell membrane preparations were solubilized in Laemmli sample buffer, fractionated on SDS–PAGE (30 μg/well) and transferred to a nitrocellulose membrane (Thermo Scientific). Immunoblots were incubated overnight at 4°C with primary antibodies including polyclonal rabbit anti-NCC (1:3,000, Millipore), rabbit anti-phospho-NCC Ser71 (pNCC S71; 1:1,000), anti-phospho-NCC Ser91 (pNCC S91; 1:1,000), anti-phospho-NCC Thr 55 (pNCC T55; 1:1,000) (generous gifts of Dario R. Alessi, University of Dundee, Dundee, UK), anti-α-ENaC (Novus Biologicals, 1:1,000), anti-β-ENaC and anti-γ-ENaC (Antikoerperonline.com, 1:1,500), anti-WNK4 (1:2,000, Novus Biologicals), and monoclonal mouse anti-β-actin (1:5,000, Sigma) in 2% (w/v) bovine serum albumin (BSA, Sigma) in a TBS-T buffer [150 mM NaCl, 10 mM Tris (pH 7.4/HCl), 0.2% (v/v) Tween-20]. After washing, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Amersham Life Sciences). Specific signal was visualized by ECL kit (Amersham Life Sciences). The protein bands were quantified by Image Quant 5.0 software (Molecular Dynamics). The expression levels were normalized to Ponceau S stain.
Andrukhova O., Slavic S., Smorodchenko A., Zeitz U., Shalhoub V., Lanske B., Pohl E.E, & Erben R.G. (2014). FGF23 regulates renal sodium handling and blood pressure. EMBO Molecular Medicine, 6(6), 744-759.
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3

Immunoprecipitation of Kidney Proteins

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Kidney cortex homogenate protein samples (1 mg) were incubated with 2 μg of anti-WNK4 (Novus Biologicals), anti-phosphoserine (Alpha Diagnostics), or anti-NCC (Millipore) antibody at 4°C overnight. The immune complexes were captured by adding 50 μl Protein A or G agarose/sepharose beads (Santa Cruz Biotechnology) and overnight incubation at 4°C with gentle rocking. The immunoprecipitates were collected by centrifugation at 1,000 × g for 5 min at 4°C and washed for four times in PBS, each time repeating the centrifugation step. After the final wash, the pellets were suspended in 40 μl of electrophoresis sample buffer and boiled for 2–3 min. Western blot analysis was performed as described above using a primary anti-NCC, anti-phosphoserine, or anti-WNK4 antibody.
Andrukhova O., Slavic S., Smorodchenko A., Zeitz U., Shalhoub V., Lanske B., Pohl E.E, & Erben R.G. (2014). FGF23 regulates renal sodium handling and blood pressure. EMBO Molecular Medicine, 6(6), 744-759.
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4

Quantification of Ubiquitylated WNK4 Protein

Cited 2 times
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Western blotting was performed as described (47 (link)). Plasma membrane fraction was purified using plasma membrane isolation kit (Invent biotechnologies). Enrichment in plasma proteins was confirmed in the laboratory (54 (link)). Equal amounts of protein were mixed with Laemmli sample buffer, separated on polyacrylamide gel, and transferred to nitrocellulose membrane. The membrane was incubated with primary and peroxidase-conjugated secondary antibodies, followed by imaging using ECL reagents (Perkin-Elmer). Tubulin and Ponceau S staining were used to ensure equal loading and transfer of samples. For the detection of ubiquitylated WNK4, WNK4-HA was purified by HA- immunoprecipitation (IP) from the cell lysates, and the ubiquitylated WNK4 was detected by Western blotting using anti-ubiquitin antibody (47 (link)). Monoclonal mouse antibody against KLHL3S433-P was created in the laboratory as described (35 (link)) and was further characterized in this study. Rabbit polyclonal antibodies against NCC phosphorylated at T53 is kindly provided by Johannes Loffing, University of Zurich, Zurich. Other antibodies include anti-KLHL3 (19 (link)), anti-FLAG, anti-tubulin (all from Sigma), anti-WNK4 (created in the laboratory) (35 (link)), anti-phospho SPAK/OSR1, anti-NCC (all from Millipore), anti-total SPAK, anti-ubiquitin, anti-WNK1 (55 (link)), and anti-calcineurin A (Cell Signaling).
Ishizawa K., Wang Q., Li J., Yamazaki O., Tamura Y., Fujigaki Y., Uchida S., Lifton R.P, & Shibata S. (2019). Calcineurin dephosphorylates Kelch-like 3, reversing phosphorylation by angiotensin II and regulating renal electrolyte handling. Proceedings of the National Academy of Sciences of the United States of America, 116(8), 3155-3160.
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