ILC2s were cultured in the presence of rm-IL-2 (10 ng/mL) and rm-IL-7 (10 ng/mL). When indicated, ILC2s were incubated with purified mouse C1q (10 μg/mL, Complement Technology) or with 15 μM of the SHP-1 inhibitor, known as Tyrosine Phosphatase Inhibitor 1 (TPI-1, MedChemExpress)39 (link),43 . The BD Biosciences Cytofix/Cytoperm kit was used and followed by intracellular staining with PE anti-p65 (clone 532301, R&D Systems), BV421 anti-AKT (pS473) (clone M89-61, BD Biosciences), PE-Cy7 anti-BCL-2 (clone 10C4, eBioscience) and unconjugated anti-SHP-1 (clone SR41-02, ThermoFisher) followed by Alexa Fluor 647 goat anti-rabbit IgG secondary antibody (Jackson ImmunoResearh). Intranuclear staining was performed using the Foxp3 Transcription Factor Staining Kit (ThermoFisher) according to the manufacturer’s instructions and APC anti-mouse Ki67 (SolA15, ThermoFisher) was used to assess proliferation. The CellTrace Violet Cell Proliferation Kit (ThermoFisher) was also used in some experiments, according to manufacturer’s instructions. In parallel, the levels of IL-5, IL-6, and IL-13 were measured in supernatants using Legendplex multiplex kits (BioLegend) and data were analyzed via the LEGENDplex data analysis software v8.0. GM-CSF was quantified using ELISA kit from BioLegend.
Tpi 1
Manufactured by MedChemExpress
Sourced in United States
TPI-1 is a laboratory instrument used for the determination of triosephosphate isomerase (TPI) enzyme activity. TPI-1 provides a convenient method for measuring the enzymatic conversion of dihydroxyacetone phosphate to D-glyceraldehyde 3-phosphate. The instrument is designed to perform this analysis accurately and efficiently in a controlled laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s modified eagle s medium dmem f12, by Thermo Fisher Scientific (3 mentions)
Glucose free dulbecco s modified eagle s medium, by Thermo Fisher Scientific (3 mentions)
Phosphate buffered saline solution pbs, by Beyotime (3 mentions)
Penicillin streptomycin pen strep, by Beyotime (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Poly l lysine (pll), by Merck Group (3 mentions)
Cck 8 kit, by Dojindo Laboratories (3 mentions)
Cytofix cytoperm kit, by R&D Systems (1 mentions)
Pe cy7 anti bcl 2 clone 10c4, by Thermo Fisher Scientific (1 mentions)
Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (1 mentions)
Legendplex multiplex kits, by BioLegend (1 mentions)
Celltrace violet cell proliferation kit, by Thermo Fisher Scientific (1 mentions)
4 protocols using tpi 1
1
ILC2 Signaling Profiling and Cytokine Analysis
2
Astrocyte Culture and Inflammation Modulation
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Adult male Sprague-Dawley (SD) rats (weighing 250–280 g) were obtained from the Animal Experimental Center of Chongqing Medical University and used for the in vivo study. Primary astrocytes were extracted from the cerebral cortices of newborn SD rats and cultured.
Glucose-free Dulbecco’s modified Eagle’s medium, Dulbecco’s modified Eagle’s medium (DMEM)/F12, and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank’s solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (NJ, USA).
Glucose-free Dulbecco’s modified Eagle’s medium, Dulbecco’s modified Eagle’s medium (DMEM)/F12, and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank’s solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (NJ, USA).
3
Astrocyte Activation and Inflammatory Responses
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Adult male Sprague-Dawley (SD) rats (between 250-280g in weight) were obtained from the Animal Experimental Center of Chongqing Medical University and used for the in vivo study. Primary astrocytes were extracted from the cerebral cortices of newborn SD rats and cultured.
Glucose-free Dulbecco's modified Eagle's medium, Dulbecco's modified Eagle's medium (DMEM)/F12 and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank's solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6 and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (New Jersey, USA)
Glucose-free Dulbecco's modified Eagle's medium, Dulbecco's modified Eagle's medium (DMEM)/F12 and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank's solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6 and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (New Jersey, USA)
4
Astrocyte Culture and Inflammatory Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adult male Sprague-Dawley (SD) rats (weighing 250-280 g) were obtained from the Animal Experimental Center of Chongqing Medical University and used for the in vivo study. Primary astrocytes were extracted from the cerebral cortices of newborn SD rats and cultured.
Glucose-free Dulbecco's modified Eagle's medium, Dulbecco's modified Eagle's medium (DMEM)/F12 and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank's solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6 and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (New Jersey, USA)
Glucose-free Dulbecco's modified Eagle's medium, Dulbecco's modified Eagle's medium (DMEM)/F12 and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Trypsin and Hank's solution were obtained from HyClone (Logan, UT, USA). Penicillin/streptomycin (Pen/Strep) and phosphate-buffered saline solution (PBS) were obtained from Beyotime (Shanghai, China). Poly-L-lysine was purchased from Sigma-Aldrich (Milan, Italy). IL-1β, IL-6 and TNF-α enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster (Wuhan, China), and a CCK-8 kit was obtained from Dojindo (Kumamoto, Japan). TPI-1 was purchased from MedChem Express (New Jersey, USA)
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