rabbit polyclonal antibody against rat p2x7  (Abcam)

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Immunohistochemical staining of the dental pulp of rat. Normal group: A1 crown pulp; A2 upper 1/3 of the root; A3 middle 1/3 of the root; and A4 root tip 1/3; NS group: B1 crown pulp; B2 upper 1/3 of the root; B3 middle 1/3 of the root; and B4 root tip 1/3; LPS group: C1 crown pulp; C2 upper 1/3 of the root; C3 middle 1/3 of the root; and C4 root tip 1/3; Transverse images of the mesial root were selected for the pulp parts, the arrows indicate the P2X7 receptor expressed in the odontoblast layer in a yellowish-brown granular form. Scale bar = 50 μm.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Western Blotting bands of the P2X7 receptor protein in the dental pulp tissue of rats in each group.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Western Blot

Journal: Molecular Pain

Article Title: Activation of the P2X7 receptor in the dental pulp tissue contributes to the pain in rats with acute pulpitis

doi: 10.1177/17448069221106844

Figure Lengend Snippet: Comparison of the P2X7 receptor protein expression in the dental pulp tissue of rats in each group. *** p < 0.001, compared to the normal group; and ▲▲▲ p < 0.001, compared to the NS group.

Article Snippet: Escherichia coli lipopolysaccharide (LPS) was purchased from Sigma, USA (L2880, 1 mg/mL, diluted in NS); Rabbit polyclonal antibody against rat P2X7 was bought from Abcam (USA); Rabbit Hypersensitivity two-step Detection Kit, bovine serum protein, and 3,3’-diaminobenzidine (DAB) color Development Kit were acquired from Beijing Zhongshan Jinqiao; P2X7 receptor antagonist A-740003 was procured from TargetMol (USA).

Techniques: Expressing

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: Both P2X7 receptor antagonists, injected at light onset (filled bars, ●), suppressed NREMS compared with saline injections (open bars, ○). oxidized ATP (OxATP) attenuated SWA during the first 12 h at the 1–4 Hz frequencies. Although EEG SWA was not significantly affected by A438079 during the first 6 h. With select 2-h time bins A438079 induced a biphasic effect, initially suppressing but then augmenting the EEG power spectra at different frequencies (see text for details, data not shown). Two smaller doses of A438079 were also tested, 1 nmol and 10 nmol; neither dose affected any of the parameters measured (*P < 0.05 between control and test days).

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques: Injection

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: Diurnal variation in the relative amount of interleukin-1β (IL1) mRNA and P2X7 receptor mRNA and protein in the hypothalamus and somatosensory cortex. A: Western blot analyses of somatosensory cortex samples showed variation of P2X7 receptor protein levels at 1500 (3 h after light onset) or 0300 (3 h after dark onset). No differences in time of day were observed in the standard, β-actin. B: time course of P2X7 receptor mRNA and protein (▴) and IL1 mRNA (●). The levels of P2X7 receptor protein (somatosensory cortex) and mRNA [(hypothalamus (□) and somatosensory cortex (▼)] were greater during the light period than during the dark period (black horizontal bar indicates dark period). Expression of protein levels was normalized to β-actin levels. *Statistical significance (P < 0.05) from values obtained at 2100; +significant differences from dark onset; #significant differences from values obtained 12 h before.

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques: Western Blot, Expressing

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: Expression of P2X7 receptor mRNA in the somatosensory cortex and hypothalamus after 6 h of sleep deprivation (SD) or 2 h and 5 h after intracerebroventricular injection of 2.5 ng of IL1. Left: SD significantly decreased the levels of P2X7 receptor mRNA in the somatosensory cortex, and IL1 enhanced P2X7 mRNA levels both 2 and 5 h after the injection. Right: in the hypothalamus SD enhanced P2X7 mRNA levels, and IL1 significantly decreased P2X7 receptor mRNA. *Significant difference from corresponding control (C).

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques: Expressing, Injection

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: IL1 enhances duration of NREMS and inhibits EEG SWA during NREMS after intraperitoneal injections in P2X7 receptor KO and WT mice. ●, Data from IL1-treated mice; ○, values from saline-treated control mice. Unlike intracerebroventricular or intravenous injections in rats, intraperitoneal injections of IL1 reduced EEG power as previously reported (37). The responses to IL1 were similar in both genotypes (WT, left; KO, right), suggesting that IL1's influence on sleep results from a step downstream from the ATP-P2 receptor interaction. *P < 0.05 difference between baseline and test days.

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques:

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: Sleep responses induced by sleep deprivation were attenuated in P2X7 receptor knockout (KO) mice compared with wild type (WT) control mice. Mice were deprived of sleep from 0600 to 1200 and then allowed to sleep ad libitum. WT mice exhibited typical NREMS rebound (right, inset) and EEG SWA enhancements (left) during the first 2 h after deprivation as previously reported (24). In contrast, P2X7 receptor KO mice had attenuated NREMS and EEG SWA responses during the same period. Both strains had reduced EEG SWA for postdeprivation hours 6–22 (1800–1000) as previously reported (24). *P < 0.05 difference between WT and KO mice for NREMS and EEG SWA. ○, Baseline values; ●, values after sleep deprivation.

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques: Knock-Out

Journal: Journal of Applied Physiology

Article Title: ATP and the purine type 2 X7 receptor affect sleep

doi: 10.1152/japplphysiol.00586.2010

Figure Lengend Snippet: Extracellular ATP involvement in sleep regulation. ATP is released into the extracellular space as a consequence of cell activity during neuro- or gliotransmission. Extracellular ATP then activates P2 receptors (R), e.g., P2X7, that in turn are involved in IL1 processing and release as well as release of other sleep regulatory substances such as tumor necrosis factor and brain-derived neurotrophic factor (not shown). IL1, in turn, activates nuclear factor-κB (NF-κB), leading to changes in receptor trafficking. This changes the cell's long-term sensitivity to neurotransmitters such as glutamate (glu) and to neuromodulators such as adenosine. Extracellular ATP is also catabolized to adenosine via the actions of ectonucleotidases such as CD39 and CD73; this action is faster than the ATP-P2-induced changes in transcription and translation. Both actions of ATP are likely involved in sleep regulation. AMPA, amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid.

Article Snippet: After centrifugation the supernatant (20 μg) was subjected to 4–20% SDS-PAGE gel (Bio-Rad, Hercules, CA), transferred to nitrocellulose membranes, and then incubated overnight with a rabbit polyclonal antibody against rat P2X7 receptor (1:15,000, Alomone Labs, Jerusalem, Israel) and 1:40,000 dilution of monoclonal mouse antibody β-actin (Sigma-Aldrich).

Techniques: Activity Assay, Derivative Assay