5′-I-RTX was purchased from Sigma Aldrich. Other reagents were sourced as described.13 (link) Functional studies were conducted on a fluorometric imaging plate reader (FLIPRTETRA) which monitored effects of capsaicin or mild acid on Ca2+ uptake into recombinant HEK293 cells expressing rat or human TRPV1 as described previously.13 (link) Antagonism of heat activation was measured at 48°C by using a custom made device (GNF Systems, San Diego, CA) for the control of temperature. Telemeters were inserted into male Sprague-Dawley rats and temperature was monitored as detailed elsewhere.13 (link) Pharmacokinetic parameters were monitored by procedures as described.13 (link)
5 irtx
Manufactured by Merck Group
5-IRTX is a reagent used in research laboratories. It is a chemical compound that can be utilized for various analytical techniques. The core function of 5-IRTX is to serve as a labeling or detection agent, but a detailed description of its intended use cannot be provided without the risk of bias or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Concanavalin a (cona), by Merck Group (2 mentions)
Fk506, by Merck Group (2 mentions)
Antimouse cd25, by Merck Group (2 mentions)
Opteia kits for il 2, by Merck Group (2 mentions)
Antimouse cd69, by Cytek Biosciences (2 mentions)
Anti mouse trpv1, by Cytek Biosciences (2 mentions)
Rpmi 1640 cell culture medium, by PAN Biotech (2 mentions)
Dmem cell culture medium, by HiMedia (2 mentions)
Antimouse cd28, by Cytek Biosciences (2 mentions)
Fetal bovine serum (fbs), by PAN Biotech (2 mentions)
Rbc lysis buffer, by HiMedia (2 mentions)
Penicillin, by HiMedia (2 mentions)
L glutamine, by HiMedia (2 mentions)
Streptomycin, by HiMedia (2 mentions)
Anti mouse cd3, by Alomone (2 mentions)
Capsaicin, by Merck Group (1 mentions)
Sb366791, by Merck Group (1 mentions)
Capsazepine, by Merck Group (1 mentions)
Anandamide, by Merck Group (1 mentions)
4 protocols using 5 irtx
1
TRPV1 Receptor Functional Assays
2
Quantifying TRPV1 Receptor Binding
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Receptor binding characteristics were studied by displacement assays using TRPV1 selective agonists (capsaicin, anandamide, and RTX; Sigma Aldrich) and antagonists (capsazepine, 5-I-RTX, and SB-366791; Sigma Aldrich). Aliquots of 100 µL of membrane suspension (1 mg protein/mL) were incubated with increasing concentrations (10-10-10-3 M) of the TRPV1 unlabelled agonists and antagonists in presence of 1.2 nM 3[H]RTX. The concentration of unlabelled ligand that it took to inhibit 50% (IC50) of 3[H]RTX specific binding was determined from the competition curves obtained by nonlinear regression analysis of the data (GraphPad Prism; GraphPad Software). The fit of the one-site model was tested by R2 by nonlinear regression and by the runs test. The affinity constant (Ki) for the competitors were calculated using the Cheng-Prussoff equation [17 (link)].
3
Murine T Cell Activation Assay
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FK506 (Cat. no. F4679-5MG), Concanavalin A (ConA) (Cat. no. C0412-5MG), 5 -IRTX (Cat. no. I9281-1MG) were purchased from Sigma Aldrich (St. Louis, MO, USA); antimouse CD25 (Cat. no. 553866), OptEIA kits for IL-2 (Cat. no.555148), , TNF (Cat. no. 558534) for sandwich ELISA were from BD Biosciences, SJ, CA, USA.; antimouse CD69 (Cat. no. 35-0691-U100), anti-mouse CD90.2 (Cat. no. 20-0903-U100) from Tonbo Biosciences, San Diego, CA, USA; anti-mouse TRPV1 (Cat. no. ACC-029) from Alomone Laboratories (Jerusalem, Israel); anti-mouse CD3 (Cat. no. 40-0032-U500) and antimouse CD28 (Cat. no. 40-0281-U500) [functional assay grade] were procured from Tonbo Biosciences, San Diego, CA, USA. Secondary anti-rabbit Alexa Fluor 488 and T cell isolation kit (Dynabeads™ Untouched™ Mouse T Cells Kit, Cat. no. 11413D) were procured from Invitrogen, Carlsbad, CA USA. RPMI-1640 cell culture medium and FBS were purchased from PAN Biotech, Aiden Bach, Germany; DMEM cell culture medium, 10X RBC lysis buffer, 10X PBS, L-glutamine, penicillin, streptomycin was from Himedia Laboratories, Mumbai, Maharashtra, India.
4
Murine T Cell Activation Assay
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FK506 (Cat. no. F4679-5MG), Concanavalin A (ConA) (Cat. no. C0412-5MG), 5 -IRTX (Cat. no. I9281-1MG) were purchased from Sigma Aldrich (St. Louis, MO, USA); antimouse CD25 (Cat. no. 553866), OptEIA kits for IL-2 (Cat. no.555148), , TNF (Cat. no. 558534) for sandwich ELISA were from BD Biosciences, SJ, CA, USA.; antimouse CD69 (Cat. no. 35-0691-U100), anti-mouse CD90.2 (Cat. no. 20-0903-U100) from Tonbo Biosciences, San Diego, CA, USA; anti-mouse TRPV1 (Cat. no. ACC-029) from Alomone Laboratories (Jerusalem, Israel); anti-mouse CD3 (Cat. no. 40-0032-U500) and antimouse CD28 (Cat. no. 40-0281-U500) [functional assay grade] were procured from Tonbo Biosciences, San Diego, CA, USA. Secondary anti-rabbit Alexa Fluor 488 and T cell isolation kit (Dynabeads™ Untouched™ Mouse T Cells Kit, Cat. no. 11413D) were procured from Invitrogen, Carlsbad, CA USA. RPMI-1640 cell culture medium and FBS were purchased from PAN Biotech, Aiden Bach, Germany; DMEM cell culture medium, 10X RBC lysis buffer, 10X PBS, L-glutamine, penicillin, streptomycin was from Himedia Laboratories, Mumbai, Maharashtra, India.
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