|Reactivity||Human, Mouse, Rat|
|Calculated MW||82026 Da|
|Homology||Rat, mouse, pig - identical.|
|Other Names||Small conductance calcium-activated potassium channel protein 3, SK3, SKCa 3, SKCa3, KCa23, KCNN3, K3|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide DTSGHFHDSGVGDLDEDPKC, corresponding to amino acid residues 2-21 of human KCa2.3 (SK3) (Accession Q9UGI6). Intracellular, N-terminus.|
|Peptide Confirmation||Confirmed by mass-spectrography and amino acid analysis.|
|Application Details||Western blot analysis (WB): - Human iPS cells (1:500) (see Linta, L. et al. (2013) in Product Citations). - Mouse and human uterine tissue (1:100) (see Pierce, S.L. and England, S.K. (2010) in Product Citations). - Rat mesenteric arteries (see Hilgers, R.H. et al. (2010) in Product Citations). Immunohistochemistry (IH): - Mouse VNO neurons, also tested in conditional KO mice (see Kim, S. et al. (2012) in Product Citations). Rat mesenteric artery parafin-embedded sections (see Hilgers, R.H. et al. (2010) in Product Citations). Immunocytochemistry (IC): - Human iPS cells (1:100) (see Linta, L. et al. (2013) in Product Citations). - CHO-K1 cells expressing human SK3 (1:1000) (see Terstappen, G.C. et al. (2001) in Product Citations).|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||25 µl, 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||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Control Antigen Reconstitution||100 µl water.|
|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.
KCa2.3 is a member of the Ca2+-activated K+ channels family with small conductance that includes KCa2.1 (SK1) and KCa2.2 (SK2). The channel is voltage insensitive and is activated by intracellular Ca2+ in the submicromolar range. It has, though, a similar topology to that of voltage-dependent K+ channels (Kv channels), that is six transmembrane domains and intracellular N- and C-termini. The functional channel of all the KCa2 family members is a multimeric protein composed of four pore-forming subunits. KCa2 channels are extremely sensitive to the levels of intracellular Ca2+ and concentrations as low as 300-700 nM can open the channels very rapidly (5-15 ms). Hence, the KCa2 channels are highly sensitive and fast Ca2+ sensors resembling other known Ca2+-binding proteins. This type of Ca2+-dependent activation is achieved by the constitutive binding of the KCa2 channels to calmodulin, a highly expressed Ca2+-binding protein via a calmodulin-binding domain situated at the cytoplasmic C-termini. Pharmacologically, the KCa2 channels are the only known targets of the bee venom toxin Apamin, with KCa2.1 being the less sensitive, KCa2.2 the most sensitive and KCa2.3 showing intermediate sensitivity. KCa2.3 is predominantly expressed in the nervous system although expression in endothelial cells, heart and liver have been described. KCa2.3 is known to be involved in the regulation of neuronal excitability. They do so mainly via a phenomenon known as afterhyperpolarization in which KCa2 channels open in response to increased intracellular Ca2+ concentrations that result from the entry of extracellular Ca2+ through voltage-dependent Ca2+ channels during action potentials. In this way, KCa2 channels effectively form a Ca2+-mediated feedback loop. KCa2.3 is involved in the control of firing rate and subsequent dopamine secretion from midbrain dopaminergic neurons. Since malfunction of these neurons is involved in several pathological disorders such as Parkinson’s disease and schizophrenia, modulators of the KCa2.3 channels have been proposed to be of therapeutic value in these diseases.
References 1. Kohler, M. et al. (1996) Science 273, 1709. 2. Xia, X.M. et al. (1998) Nature 395, 503. 3. Stocker, M. (2004) Nat. Rev. Neurosci. 5, 758.
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