|Calculated MW||90855 Da|
|Homology||Mouse - identical; human, rabbit, guinea pig - 69/70 amino acid residues identical; Xenopus laevis - 61/70 amino acid residues identical. Rat CLC-4 - 46/70 amino acid residues identical; rat CLC-5 - 49/70 amino acid residues identical.|
|Other Names||H(+)/Cl(-) exchange transporter 3, Chloride channel protein 3, ClC-3, Chloride transporter ClC-3, Clcn3|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||GST fusion protein with amino acid residues 592-661 of rat CLC-3 (Accession P51792). Intracellular, near the C-terminus.|
|Peptide Confirmation||Confirmed by DNA sequence and SDSPAGE.|
|Application Details||Western blot analysis (WB): - tsA cells expressing CLC-3 (see Farmer, L.M. et al. (2013) in Product Citations). - Human glioma lysates (1:500), (see Cuddapah, V.A. et al. (2013) in Product Citations). Immunoprecipitation (IP): - tsA cells expressing CLC-3 (see Farmer, L.M. et al. (2013) in Product Citations). Immunohistochemistry (IH): - Rat brain sections (1:100) (see Farmer, L.M. et al. (2013) in Product Citations). Immunocytochemistry (IC): - tsA cells expressing CLC-3 (see Farmer, L.M. et al. (2013) in Product Citations).|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 mg/ml.|
|Buffer After Reconstitution||Phosphate buffered saline (PBS), pH 7.4, 1 % BSA, 5% sucrose, 0.025% NaN3.|
|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 PBS.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||3 µg fusion protein per 1 µg antibody.|
|Formulation||Lyophilized powder. Reconstituted anitbody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 5% sucrose, 0.025% NaN3.|
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Provided below are standard protocols that you may find useful for product applications.
CLC-3 is a member of the voltage-dependent Cl- channel (CLC) family that includes nine known members in mammals. CLC channels can be classified as plasma membrane channels and intracellular organelle channels. The first group includes the CLC-1, CLC-2 CLC-Ka and CLCKb channels. The second group comprises the CLC-3, CLC-4, CLC-5, CLC-6 and CLC-7. CLC channels that function in the plasma membrane are involved in the stabilization of membrane potential and in transepithelial transport. The presumed function of the intracellular CLC channels is support of the acidification of the intraorganellar compartment. In this regard, recent reports indicate that ClC-4 and ClC-5 (and by inference ClC-3) can function as Cl-/H+ antiporters.1, 2 The functional unit of the CLC channels is a dimer with each subunit forming a proper pore. Although the crystal structure of bacterial CLC channels was resolved, the topology of the CLC channels is complex and has not been fully elucidated. It is generally accepted that both the N- and C- terminus domains are intracellular while the number and configuration of the transmembrane domains vary greatly between different models. 1,2 CLC-3 is widely distributed with prominent expression in tissues of neuroectoderm origin. In the brain, it is highly expressed in the hippocampus, olfactory bulb and olfactory cortex. The channel is also prominently expressed in aortic and coronary vascular smooth muscle cells, aortic endothelial cells and tracheal and alveolar epithelial cells. The physiological function of CLC-3 is not entirely clear, but it has been suggested that CLC-3 generates a shunt current of chloride for v-H+-ATPases, thereby aiding the acidification of endosomes and synaptic vesicles as well as lysosomes. Disruption of the ClC-3 gene in mice causes severe neuronal loss, leading to a complete loss of the hippocampus in adult mice. In addition, CLC-3 has been shown to have a critical role in the respiratory burst and phagocytosis of polymorphonuclear cells, a key cell type of innate host defense. 3,4
References 1. Jentsch, T.J. et al. (2002) Physiol. Rev. 82, 503. 2. Babini, E. and Pusch, M. (2004) Physiology 19, 293. 3. Stobrawa, S.M. et al. (2001) Neuron 29, 185. 4. Moreland, J.G. et al. (2006) J. Biol. Chem. 281, 12277.
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