A total of 5 × 104 CAR T cells were co-cultured with 5 ×104 tumour cells in 200 μl of complete T cell medium (AIM-V or RPMI) without IL-2 in a 96-well plate, all in triplicate. Twenty-four hours after co-culture, culture supernatants were collected, diluted 20- to 100-fold and analysed for IL-2 and IFNγ using ELISA MAX kits (BioLegend) and Nunc Maxisorp 96-well ELISA plates (Thermo Fisher Scientific). Absorbance readings were collected on a Tecan Spark plate reader or a BioTek Synergy H1 running Gen5 v.2.00.18. For FOXO1i assays, the co-culture medium included concentrations of AS1842856 that were used during T cell expansion.
Spark plate reader
Manufactured by Agilent Technologies
Sourced in United States
The Spark plate reader is a versatile instrument designed for various types of quantitative and qualitative biochemical and cell-based assays. It can measure absorbance, fluorescence, and luminescence signals in a 96- or 384-well microplate format. The Spark plate reader provides accurate and reproducible results, making it a reliable tool for applications such as enzymatic activity analysis, protein quantification, and cell viability assessment.
Automatically generated - may contain errors
Lab products found in correlation
Elisa max kit, by BioLegend (1 mentions)
Nunc maxisorp 96 well elisa plate, by Thermo Fisher Scientific (1 mentions)
Synergy h1, by Agilent Technologies (1 mentions)
Hydrochloric acid (hcl), by Chempur (1 mentions)
Acetic acid, by Chempur (1 mentions)
Trichloroacetic acid, by Chempur (1 mentions)
Methanol, by Chempur (1 mentions)
Bodipy 581 591 c11, by Thermo Fisher Scientific (1 mentions)
2 4 6 tris 2 pyridyl s triazine, by Merck Group (1 mentions)
Thiobarbituric acid tba, by Serva Electrophoresis (1 mentions)
Ferric chloride, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
2 protocols using spark plate reader
1
Quantifying CAR T-cell Cytokine Secretion
2
Antioxidant Assay Protocols
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Bodipy™ 581/591 C11 was obtained from ThermoFisher. 4',5,7-Trihydroxy-6-methoxyflavone (hispidulin) was purchased from Medchemexpress. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 2,2'-azino-bis(3-ethylobenzthiazoline-6sulfonic acid, diammonium salt) (ABTS), 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB), potassium persulfate, sodium ethylenediaminetetraacetate (EDTA) and ferric chloride were from Merck (Poznań, Poland). Phosphate-buffered saline (PBS), sodium dodecyl sulfate (SDS), monosodium phosphate, disodium phosphate and dimethylsulfoxide (DMSO) were from Bioshop (Burlington, ON, Canada).
Thiobarbituric acid (TBA) was from Serva Electrophoresis GmbH (Heidelberg, Germany). Methanol, acetic acid, hydrochloric acid and trichloroacetic acid were from Chempur (Piekary Śląskie, Poland).
Sodium acetate was from POCh (Gliwice, Poland).
Transparent and black flat-bottom 96-well plates (Greiner, Kremsmünster, Austria) were used for the assays. Absorbance and fluorescence was measured in a Spark plate reader (BioTek, Winooski, VT, USA).
Thiobarbituric acid (TBA) was from Serva Electrophoresis GmbH (Heidelberg, Germany). Methanol, acetic acid, hydrochloric acid and trichloroacetic acid were from Chempur (Piekary Śląskie, Poland).
Sodium acetate was from POCh (Gliwice, Poland).
Transparent and black flat-bottom 96-well plates (Greiner, Kremsmünster, Austria) were used for the assays. Absorbance and fluorescence was measured in a Spark plate reader (BioTek, Winooski, VT, USA).
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