5′-N-ethylcarboxamidoadenosine (NECA) and 4-[2-[[6-Amino-9-(N-ethyl-β-D-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid (CGS 21680) were purchased from Sigma-Aldrich (St. Louis, MO, USA). NECA and CGS-21680 were prepared as 10 mM stock solutions using DMSO (Sigma, St. Louis, MO) followed by serial dilutions with 50% DMSO and distilled water and a further dilution to the desired concentration was achieved with distilled water (final DMSO concentration of <1%) [9 (link)]. Anti-A2AAR antibody was purchased from EMD Millipore. Alexa 533-conjugated goat anti-mouse secondary antibody was purchased from Invitrogen, and DRAQ5 was purchased from Abcam.
Cgs 21680
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
The CGS 21680 is a laboratory equipment product manufactured by Merck Group. It serves as a tool for researchers and scientists, but a detailed factual description of its core function cannot be provided while maintaining an unbiased and concise approach. Further information may be available from the manufacturer.
Automatically generated - may contain errors
Lab products found in correlation
Sch58261, by Merck Group (6 mentions)
Zm241385, by Merck Group (4 mentions)
Dpcpx, by Merck Group (3 mentions)
Zm241385, by Bio-Techne (3 mentions)
Mrs1754, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Apyrase, by Merck Group (2 mentions)
Atpγs, by Merck Group (2 mentions)
Pom 1, by Bio-Techne (2 mentions)
Recombinant il 23, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Golgiplug, by BD (2 mentions)
Bay60 6583, by Merck Group (2 mentions)
Caffeine, by Merck Group (2 mentions)
Adenosine, by Merck Group (2 mentions)
Quinpirole hcl, by Merck Group (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Euroclone (2 mentions)
33 protocols using cgs 21680
1
Pharmacological Manipulation of A2A Receptors
2
Adenosine Receptor Antagonist Assay
3
Modulation of Intestinal and Spleen Cell Responses
4
Isolation and Culture of Intestinal and Spleen Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intestinal LPLs or spleen cells were isolated from Rag1−/− mice and cultured in completed RPMI 1640 medium supplemented with 10% FBS, 2‐Mercaptoethanol (50 μmol L−1, Gibco, Waltham, MA, USA), l ‐glutamine (2 mmol L−1, Gibco) and antibiotics (Penicillin and Streptomycin, 100 U mL−1, Gibco). When indicated, recombinant IL‐23 (20 ng mL−1, eBioscience, San Diego, CA, USA), POM‐1 (2–50 μmol L−1, Tocris Bioscience), APCP (2–50 μmol L−1), Apyrase (10 μmol L−1, Sigma), ATPγS (5–10 μmol L−1, Sigma), and A2A agonist CGS 21680 (1–10 μmol L−1, Merck Chemicals, Darmstadt, Germany) and their respective vehicle control (i.e. DMSO or dH2O) or combination of these reagents were added into cell cultures. For detecting cytokines in supernatants, cells were cultured for overnight, and for intracellular staining of IL‐22, cells were stimulated for 4 h in the presence of GolgiPlug (BD Bioscience, San Jose, CA, USA). In vitro cell cultures were performed in triplicate and repeated two or more times.
5
Purinergic signaling modulators' effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NECA and withaferin A were purchased from Biomol (Hamburg, Germany). MRS1754, SCH58261, BAY60-6583 and Lucifer Yellow (LY) were purchased from Sigma-Aldrich (Taufkirchen, Germany). CGS21680 was from Merck Millipore (Darmstadt, Germany). SQ22536 was from Enzo Life Sciences (Lörrach, Germany). Adenosine-3',5'-cyclic monophosphorothioate (Rp-cAMPS) was from Biolog Life Science Institute (Bremen, Germany). NECA, withaferin A, MRS1754, SCH58261, CGS21680, and BAY60-6583 were dissolved in DMSO while Rp-cAMPS and SQ22536 were dissolved in water. All inhibitors were preincubated for 30 min prior to addition of NECA. The vehicles DMSO or water were added to control cells in all experiments at maximal concentrations of 0.1% or 0.3%, respectively.
6
Isolation and Culture of Primary Human and Murine Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary Human Umbilical Vein Endothelial Cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured in endothelial cell growth medium (EGM, Promocell) to passage 4 or 5 before being treated with various CpG-ODNs (Supplementary Table S1 ) (TIB Molbiol, Berlin, Germany). Pericytes were immuno-sorted from saphenous vein leftovers of coronary artery bypass graft surgery patients as described previously61 (link)62 (link). Bone marrow derived macrophages (BMDM) were prepared from 4–6 weeks old C57BL/6 mice as described before63 (link). BMDM cultures were stimulated with IL-4 (20 U/mL) (PeproTech, UK), LPS (1 μg/mL) (Sigma) and CGS-21680 (10 nM) (Sigma). Studies on cells from human subjects complied with the principles stated in the “Declaration of Helsinki” and were covered by approval (06/Q2001/197) from the Bath Research Ethics Committee.
7
Characterization of Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isoproterenol, formoterol, GO6983, GO6976, RO32–0432, GF109203X, H89, PSB603, ESI-09, TPPB, NECA (adenosine-5′-N-ethyluronamide), MRS5698 and MRS2365 ([[(1R,2R,3S,4R,5S)-4-[6-amino-2-(methylthio)-9H-purin-9-yl]-2,3-dihydroxybicyclo[3.1.0]hex-1-yl]methyl] diphosphoric acid monoester trisodium salt) were purchased from Tocris (Ellisville, MO). CGS21680">CGS21680 , carbachol, cholera toxin (CTX) and PTX were from Sigma (St. Louis, MO). YM254890, enzastaurin, LY333531, VTX-27 and AB928 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). BAY60-6583 (LUF6210, termed hereafter ‘BAY’) was synthesized at Leiden/Amsterdam Center for Drug Research and was provided by Ad IJzerman (Leiden, The Netherlands). All compounds were dissolved in DMSO except that CTX and PTX were in water, and proper controls were included in all experiments. AlphaScreen cAMP kit, SureFire p-ERK1/2 (Thr202/Tyr204) Assay Kit and AlphaScreen SureFire p-Akt 1/2/3 (p-Ser473) Assay Kit were purchased from PerkinElmer (Waltham, MA). Gs-null and Gq/11-null HEK293 cells were generated at Tohoku University, Sendai, Japan. HEK293 human embryonic kidney, PC-3 human prostate cancer, NIH-3T3 mouse fibroblast, and H9C2 rat cardiomyoblast cells were from ATCC (Manassas, VA); all other reagents were from standard commercial sources and of analytical grade.
8
Vascular Reactivity Pharmacology Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phenylephrine, acetylcholine (ACh) (Sigma Chemicals, St. Louis, MO), and angiotensin II (Human), BACHEM (Bubendorf, Switzerland) dissolved in distilled water. N-(Methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MS-PPOH), dibromo-dodecenyl-methylsulfimide (DDMS) (Cayman Chemicals), 5'-N-ethylcarboxamidoadenosine (NECA), and CGS 21680 (Sigma Chemicals, St. Louis, MO) were dissolved in dimethyl sulfoxide (DMSO) (Nayeem et al., 2013 (link)).
9
Caffeine-Induced Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Caffeine was purchased from Sigma Chemical Co (St. Louis, MO, USA). Acetaldehyde was purchased from Tianjin DaMao Chemical Reagent. Dimethyl sulfoxide (DMSO) was purchased from Sigma Inc. CGS21680(A2AR agonist), ZM241385(A2AR antagonist) and SB202190(P38 MAPK inhibitor) were purchased from Tocris (Ellisville, MO, USA). U0126 (MEK1/2 inhibitors), antibodies against SRC, ERK1/2, P38 MAPK, and phosphorylation-dependent antibodies against SRC, ERK1/2, P38 MAPK were purchased from Cell Signaling Technology (Beverly, MA, USA). H89 (PKA inhibitor), PP2 (SRC inhibitor) were purchased from Santa Cruz (CA, USA). Secondary antibodies including goat anti-rabbit immunoglobulin G (IgG) horse radish peroxidase (HRP) and goat anti-mouse IgG HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, California, USA). RevertAidTM First Strand cDNA Synthesis Kit and Brilliant SYBR Green QPCR Master were purchased from Fermentas (CA, USA). 125I-cAMP radioimmunoassay kit was purchased from Shanghai University of Traditional Chinese Medicine (Shanghai, China). All the other chemicals used were of the highest grade available commercially.
10
TrkB and Adenosine Receptor Agonist/Antagonist Preparation
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!