Isoprenaline (ISO, Sigma) was dissolved in sterile PBS and administered by continuous infusion using implantable mini-osmotic pumps (Alzet) at a dose of 30 mg/kg body mass/day. Pumps were filled and primed in a sterile environment and were inserted into the dorsal subcutaneous space caudal to the scapulae.
Isoprenaline iso
Manufactured by Merck Group
Sourced in United States
Isoprenaline (ISO) is a laboratory reagent used for various research and analytical applications. It functions as a non-selective beta-adrenergic agonist, stimulating both beta-1 and beta-2 adrenergic receptors. ISO is commonly utilized in cell-based assays, biochemical experiments, and pharmacological studies.
Automatically generated - may contain errors
Lab products found in correlation
Neurobasal a medium, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
6 well plate, by BD (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Dasatinib, by Merck Group (1 mentions)
Ici 118 551, by Merck Group (1 mentions)
Antimouse and antirabbit secondary antibodies, by Fortis Life Sciences (1 mentions)
Propranolol, by Merck Group (1 mentions)
Gsdmd, by Proteintech (1 mentions)
Il 1β, by Proteintech (1 mentions)
Aav9 nc, by Genechem (1 mentions)
Collagen 3, by Proteintech (1 mentions)
Tgf β1 100 21, by Thermo Fisher Scientific (1 mentions)
Sirt1, by Proteintech (1 mentions)
Caspase 1, by Proteintech (1 mentions)
Antibody against α sma, by Abcam (1 mentions)
Collagen 1, by Proteintech (1 mentions)
β actin, by Proteintech (1 mentions)
Nlrp3, by Proteintech (1 mentions)
9 protocols using isoprenaline iso
1
Isoprenaline Infusion for Research
2
Pineal Gland Organotypic Culture Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Organotypic cultures were prepared from 16-day-old rats. Pineal glands were removed after rapid decapitation and placed onto cell culture inserts with a pore size of 0.001 mm (BD Falcon, Tewksbury, MA, USA) in 6-well plates (Falcon). Inserts were submerged in 1 mL of Neurobasal A medium supplemented with 2% serum-free B-27, 50 U/mL penicillin, 50 µg/mL streptomycin, and 0.5 mM L-glutamine (Thermo Fisher Scientific, Waltham, MA); saturated with 95% air and 5% CO2 mixture; and incubated in a humidified 5% CO2 atmosphere at 37 °C. Organotypic cultures were allowed to stabilize, and after 3 days of cultivation, they were stimulated either with WP1066 inhibitor (WP, 5 µM) for 6 h, LPS (10 µg/mL of media) for 5 h, isoprenaline (ISO, Sigma-Aldrich (St. Louis, MO, USA), I6504; 2 µM) for 4 h, or their mutual combinations. Organotypic cultures were then frozen and processed by AA-NAT enzymatic activity assay.
3
Isoprenaline Infusion for Research
4
Cardiac Mechanics: Isometric Force Clamping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To set the feedback control parameters, a rat ventricular myocyte was electrically stimulated and stretched until a minimum force development of 0.3–0.5 μN was recorded. The forces at 10 and 30% of the developed force (i.e. of the difference between the maximum and minimum forces registered from isometric contractions) were used to designate the initial pre-load and after-load values, respectively. With these initial pre-load and after-load values set, we engaged the force clamping algorithm. When control of the cardiac cycle appeared stable, the actual protocol was started (see Supplementary material online, FigureS3, for an example of a stable recording). At each pre-load condition, the after-load was varied using a ramp function that would rise over several electrical stimulations by 0.75 µN and then decline back by 0.45 µN. The pre-load was then raised by 0.3 µN and, thereafter, by another 0.3 µN (see Supplementary material online, FigureS4 ). This protocol was repeated for 1, 2, 4, 6, and 8 Hz pacing frequencies in the presence of Tyrode or Tyrode with 100 nmol/L isoprenaline (ISO, Sigma Aldrich). More than 90% of the attempts to attach a cell was successful. Because of the elaborate protocol, the full protocol could be completed in 3–5 cells per experimental day.
5
Pharmacological Modulation of Src Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Propranolol, ICI-118,551, E, and NE, purchased from Sigma-Aldrich (St Louis, MO, USA), were dissolved in distilled water (DW) at 100 mM. Isoprenaline (ISO) and dasatinib were also purchased from Sigma-Aldrich and reconstituted in dimethyl sulfoxide (DMSO; Amresco, Solon, OH, USA) at 100 mM and at 4 mM, respectively. The primary antibody against phospho-Src (Y416) was purchased from Cell Signaling Technology (Beverly, MA, USA). Primary antibodies against Src and actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-mouse and anti-rabbit secondary antibodies were purchased from Bethyl Laboratories (Montgomery, TX, USA) and Enzo Life Sciences (Farmingdale, NY, USA), respectively.
6
Murine Cardiac Function Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isoflurane was reduced to 1.5% and animals were allowed to reach steady state conditions as indicated by constant heart rate and stroke volume, which was observed within 3–5 min. If the stabilization period extended 10 min, animals were not included into the analysis. The first two cycles of recording were performed while infusing 15 μl/min 0.9% NaCl. PV loops were recorded during two cycles of inferior caval vein (ICV) occlusion for each Isoprenaline (ISO) dose, after a new steady state was reached. To maintain constant infusion rates of 15 μl/min, increasing doses of 0.2475, 0.825, 2.475 and 8.25 ng/min Isoprenaline (ISO, Sigma Life Science) dissolved in 0.9% NaCl were infused by switching syringes. As the respiratory status has a significant impact on murine cardiac function during PV recordings (Figure S2 ), ventilation was suspended strictly at the end-expiration time point. Preload dependent parameters were recorded during the first 2 s of a measurement (Figure S3 ). To obtain preload-independent parameters, the suture around the ICV was gently lifted for the following 2 s of each measurement (Figure S3 ).
7
Profibrotic Pathway Regulation by miR-217
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isoprenaline (ISO) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in phosphate-buffered saline (PBS). TGF-β1 (100-21) was obtained from Peprotech (Rocky Hill, NJ, USA). mir-217 mimics and mir-217 inhibitor were purchased from Hanbio Biotechnology Co., Ltd. (Shanghai, China). Antibody against α-SMA was purchased from Abcam (Cambridge, MA, USA). The following antibodies were purchased from Proteintech (Chicago, IL, USA): SIRT1, collagen I, collagen III, GSDMD, Caspase-1, IL-1β, NLRP3 and β-actin. AAV9-GAS5 (recombinant adeno-associated virus expressing growth arrest specific transcript-5) and AAV9-NC were constructed by Genechem (Shanghai, China).
8
Zebrafish Heart Resection and Cardiac Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Larvae of the wild-type Turku strain of zebrafish [18 (link),19 (link)] were maintained and bred according to Westerfield [20] at the Biomedicum Helsinki Zebrafish Unit. 2–4 days post fertilization (dpf) zebrafish were used for the experiments. Zebrafish larvae were maintained in E3 medium (5.0 mM NaCl, 0.4 mM CaCl2, 0.3 mM MgSO4 and 0.2 mM KCl) and treated with isoprenaline (Iso) (Sigma-Aldrich, St. Louis, MO) when desired. For heart resection, zebrafish were anesthetized in 0.03% tricaine. Zebrafish without heart resection were euthanized in 0.03% tricaine followed by addition of ice. Experiments on zebrafish were performed in accordance with protocols approved by the National Animal Experiment Board of Finland (ESAVI/4131/04.10.07/2017). Vezf1 splice-blocking morpholino antisense oligonucleotides (SBMO) 5´-CATTGGCCTGCTGGATGGAGAAAGA-3´, Vezf1 translation-blocking MO (TBMO) 5´-ATGAACTCCAGCTCGGCTCCATTGC-3´ and random control oligo (control) 25-N were from Gene Tools, LLC (Philomath, OR). Dosage was determined by titration; 3–4 ng for SBMO and 5–7 ng for TBMO. After confirming similar findings with both morpholinos in phenotype as well as in cardiac physiology, experiments were done with SBMOs, as they better allowed simultaneous qPCR experiments. For the rescue experiments 250 pg of capped Vezf1 mRNA was co-injected with SBMO.
9
Chondrocyte cell line CHON-001 culture and treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human chondrocyte cell line CHON-001 was purchased directly from American type culture collection (ATCC, CRL-2846). Briefly, CHON-001 cells were thawed and cultured immediately in a humid incubator (5% CO2, 37°C) after receipt according to the instructions. The cells were cultured and maintained in DMEM media (Hyclone, Logan, UT, United States) containing 10% FBS and 1% penicillin/streptomycin. When cells reached 80% confluence, they were subcultured at a 1:4 ratio. Cells were collected from the petri plate for subculturing by using 2–3 ml 0.05% Trypsin-0.53 mm EDTA (Cat # 25200056) (Hyclone, Logan, UT, United States). All the experiments in this study is performed after three passages. Cells were cryopreserved with Fetal Bovine Serum (FBS) (Hyclone, Logan, UT, United States) supplemented with 10% DMSO in liquid nitrogen.
In vitro experimental design consisted of 4 groups. For each group, 1 × 105 chondrocyte cells were seeded in 48-well plate in duplicate. Group 1 was the control group without any treatment for the cells. Group 2 was treated with IL-1β (10 ng/ml) for 24 h. Group 3 was treated with β2-AR agonist Isoprenaline (ISO) (10−5 mol/L) purchased from Sigma (St. Louis, MO, United States) for 24 h. Group 4 was treated with IL-1β for 6 h and then ISO was administered up to 24 h in total. Cells were collected to perform the further experiments.
In vitro experimental design consisted of 4 groups. For each group, 1 × 105 chondrocyte cells were seeded in 48-well plate in duplicate. Group 1 was the control group without any treatment for the cells. Group 2 was treated with IL-1β (10 ng/ml) for 24 h. Group 3 was treated with β2-AR agonist Isoprenaline (ISO) (10−5 mol/L) purchased from Sigma (St. Louis, MO, United States) for 24 h. Group 4 was treated with IL-1β for 6 h and then ISO was administered up to 24 h in total. Cells were collected to perform the further experiments.
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!