|Application ||WB, IP|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||43501 Da|
|Homology||Mouse - identical; human - 15/19 amino acid residues identical; chicken - 11/12 amino acid residues identical; Xenopus - 11/11 amino acid residues identical.|
|Other Names||P2X purinoceptor 4, P2X4, ATP receptor, Purinergic receptor, P2rx4|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)KKYKYVEDYEQGLSGEMNQ, corresponding to amino acid residues 370-388 of rat P2X4 Receptorֲ (Accession P51577). Intracellular, C-terminus.|
|Peptide Confirmation||Confirmed by amino acid analysis.|
|Application Details||Western blot analysis (WB): - Mouse bone marrow-derived dendritic cells (BMDCs) (1:300) (see Sakaki, H. et al. (2013) in Product Citations). - Rat spinal cord lysate (see Lu, W.H. et al. (2013) in Product Citations). - HEK-293 cells expressing rat P2X4 receptor (see Rokic, M.B. et al. (2013) in Product Citations). Immunoprecipitation (IP): - Rat hypothalamus lysate (see Jo, Y.H. et al. (2011) in Product Citations). Immunohistochemistry (IH): - Primate retina (1:1000) (see Gu, B.J. et al. (2013) in Product Citations). - Rat spinal cord (see Lu, W.H. et al. (2013) in Product Citations). - Mouse brain (1:200) (see Jo, Y.H. et al. (2011) in Product Citations). Immunocytochemistry (IC): - Rat primary microglia (see Lu, W.H. et al. (2013) in Product Citations). - Human mesangial cells (HMC) (1:200) (see Solini, A. et al. (2007) in Product Citations). Indirect flow cytometry (IFC): - NR8383 rat alveolar macrophage cell line (1:100) (see Stokes, L. and Surprenant, A. (2009) 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||0.8 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.
The P2X receptors belong to the ligand-gated ion channel family and are activated by extracellular ATP. The structure and function of the P2X receptors, which were mainly investigated using in vitro models, indicate their involvement in synaptic communication, cell death, and differentiation. Seven mammalian P2X receptor subtypes (P2X1–P2X7) have been identified and cloned.1,2,3 All P2X receptor subtypes share the same structure of intracellular N and C-termini, two membrane-spanning domains and a large extracellular loop. All P2X subunits can assemble to form homomeric or heteromeric functional channels with the exception of P2X6, which only appears to function as part of a heteromeric complex.4-9 The various P2X receptors show distinct expression patterns. P2X1-6 have been found in the central and peripheral nervous system, while the P2X7 receptor is predominantly found in cells of the immune system.4 The P2X2 receptor subunit has a widespread tissue distribution in autonomic neurons, but it is generally found to be co-expressed with one or more subtypes. Overexpression of P2X4 was demonstrated in microglia and in the spinal dorsal horn following peripheral nerve injury. It has been suggested that activation of P2X4 along with p38 MAPK is essential for the development of allodynia (pain from a stimulus that doesn't normally elicit pain) following nerve injury. Inhibition of P2X4 expression in spinal microglia has been suggested as a novel therapeutic approach for the treatment of allodynia.10
References 1. Prasad, M. et al. (2001) J. Physiol. 573, 667. 2. Florenzano, F. et al. (2002) Neuroscience. 115, 425. 3. Ashcroft, F.M. et al. (2000) Ion Channels and Disease Ed 1, p. 405, Academic Press, San Diego. 4. Khakh, B.S. et al. (2001) Pharmacol Rev. 53, 107. 5. Ding, Y. et al. (2000) J. Auton. Nerv. Syst. 81, 289. 6. Le, K.T. et al. (1998) J. Neurosci. 18, 7152. 7. Robertson, S.J. et al. (2001) Curr. Opin. Neurobiol. 11, 378. 8. Dunn, P.M. et al. (2001) Prog. Neurobiol. 65, 107. 9. Kim, M. et al. (2001) EMBO J. 20, 6347. 10. Inoue, K. et al. (2004) J. Pharmacol. Sci. 94, 112.
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