|Calculated MW||94405 Da|
|Homology||Mouse, human - identical.|
|Other Names||Two pore calcium channel protein 1, Voltage-dependent calcium channel protein TPC1, Tpcn1, Tpc1|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)RNLRQIFQSLPPFMD, corresponding to amino acid residues 221-235 of ֲ rat Two Pore Calcium Channel Protein 1ֲ ֲ (Accession Q9WTN5). 2nd extracellular loop for Two pore calcium channel protein 1 expressed on the plasma membrane. Luminal for Two pore calcium channel protein 1 expressed on intracellular organelles.|
|Peptide Confirmation||Confirmed by 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||0.9 mg/ml.|
|Buffer After Reconstitution||Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% 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 water.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||1 µg peptide per 1 µg antibody.|
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Provided below are standard protocols that you may find useful for product applications.
Among various vertebrate species, three genes are known to encode two-pore segment channels (TPCs) termed TPC1-3. Interestingly TPC3 seems to be absent from the genomes of primates and rodents1. The primary sequence of these channels indicates the presence of two putative pore-forming repeats. Each repeat contains six transmembrane domains and a pore loop, a structure strikingly reminiscent of many voltage-gated Na+ (Nav) and Ca2+ (Cav) channels2. These twelve transmembrane structures are further thought to form functional dimers3. Both TPC1 and TPC2 show ubiquitous expression, while that of TPC1 is exceptionally high in spleen, lung, liver, and kidney2. Ca2+-mobilizing messengers such as inositol triphosphate, cyclic ADP ribose and nicotinic acid adenosine dinucleotide phosphate (NAADP) are responsible for the intracellular changes in Ca2+ ion concentration4. In contrast to the other Ca2+-mobilizing agents, NAADP, the most potent of these Ca2+ releasing molecules increases the cytosolic Ca2+ concentration via Ca2+ channels located on acidic vesicles (endolysosomes)5,6. Only quite recently, after almost a decade of being cloned, TPC1 and TPC2 were both found to be responsible for the NAADP-induced release of Ca2+ 7,8. Evidence that these two channels are indeed responsible for the release of Ca2+ is quite compelling since overexpression of TPC1 and its knockdown or point mutation of a critical residue increase and exacerbate Ca2+ release respectively7. In addition, b-cells from TPC2 knockout mice exhibited no Ca2+ release from endolysosomes upon NAADP stimulation8. Finally, in a study using immunopurified channels, it was demonstrated that TPC1 and TPC2 both respond to very low concentrations of NAADP and are unequivocally responsible for the release of Ca2+, whereas TPC3 may negatively regulate the release of Ca2+ 9. As these channels have only recently been discovered, very little is known about their physiology and gating mechanisms. Their probable involvement in a number of diseases such as lysosomal storage disease (LSDs), caused by the dysfunction of lysosomal associated proteins, has yet to be deciphered1. Abgent is pleased to offer a highly specific antibody directed against an extracellular epitope located of rat Two Pore Calcium Channel Protein 1. Anti-Two Pore Calcium Channel Protein 1 (extracellular) antibody (#AG1332) can be used in western blot and immunohistochemical applications, and has been designed to recognize Two Pore Calcium Channel Protein 1 from rat, mouse and human samples.
References 1. Zhu, M.X. et al. (2010) Am. J. Physiol. 298, C430. 2. Ishibashi, K. et al. (2000) Biochem. Biophys. Res. Commun. 270, 370. 3. Galione, A. et al. (2009) Pflugers Arch. 458, 869. 4. Berridge, G. et al. (2000) Nat. Rev. Mol. Cell Biol. 1, 11. 5. Lee, H.C. (2005) J. Biol. Chem. 280, 33693. 6. Churchill, G.C. et al. (2002) Cell 111, 703. 7. Brailoiu, E. et al. (2009) J. Cell. Biol. 186, 201. 8. Calcraft, P.J. et al. (2009) Nature 459, 596. 9. Ruas, M. et al. (2010) Curr. Biol. 20, 703.
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