|Application ||WB, LCI|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||131447 Da|
|Homology||Rat - identical; mouse - 13/14 amino acid residues identical.|
|Other Names||Solute carrier family 12 member 2, Basolateral Na-K-Cl symporter, Bumetanide-sensitive sodium-(potassium)-chloride cotransporter 1, SLC12A2, NKCC1|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)RRQAMKEMSIDQAK, corresponding to amino acid residues 828-841 of human NKCC1 (Accession P55011). 6th extracellular loop.|
|Peptide Confirmation||Confirmed by mass-spectrography and amino acid analysis.|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.6 mg/ml.|
|Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).|
|Control Antigen Storage Before Reconstitution|
|Control Antigen Reconstitution||100 µl DDW.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||1 µg peptide per 1 µg antibody.|
|Formulation||Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.|
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Provided below are standard protocols that you may find useful for product applications.
The Na-K-Cl cotransporters are a class of membrane proteins that transport Na, K, and Cl ions into and out of cells in an electrically neutral manner, in most cases with a stoichiometry of 1Na+:1K+:2Cl-. Na-K-Cl cotransporters are present in a wide variety of cells and tissues, including reabsorptive and secretory epithelia, nerve and muscle cells, endothelial cells, fibroblasts, and blood cells. Na-K-Cl cotransport plays a vital role in renal salt reabsorption and in salt secretion by intestinal, airway, salivary gland, and other secretory epithelia1. Na-K-Cl cotransport activity is inhibited by "loop" diuretics, including the clinically efficacious agents, bumetanide and furosemide. The regulation of Na-K-Cl cotransport is mediated, at least in some cases, through direct phosphorylation of the cotransport protein2. NKCC1 is one of two Na-K-Cl cotransporter isoforms that have been identified. NKCC1 is present in a wide variety of secretory epithelia and non-epithelial cells3. NKCC1 is expressed in virtually all mammalian cells and is thought to play a housekeeping role in cell volume homeostasis and the common control of cytosolic ion content. NKCC1 does not use ATP but operates using electrochemical gradient for Na+ and K+ produced by Na+-K+-ATPase4. There is considerable evidence that uptake of Cl− in immature neurons is mediated by NKCC1. High expression of NKCC1 in immature neurons plays an important role in maintaining high intracellular Cl−5.
References 1. Haas M. (1994) Am. J. Physiol. 267, C869. 2. Delpire E. et al. (1999) Nat. Genet. 22, 192. 3. Haas M. and Forbush, B. 3rd (2000) Annu. Rev. Physiol. 62, 515. 4. Ben-Ari Y. et al. (2007) Physiol. Rev. 87, 1215. 5. Delpire E. et al. (2000) News Physiol. Sci. 15, 309.
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