Human cxcl1 gro alpha quantikine elisa kit

ELISAs were performed to measure cytokine release from the IEC monolayers in both the apical and basal compartment supernatants. IEC monolayers were infected with G. intestinalis trophozoites preconditioned with DMEM/F-12 or DMEM/F-12+FBS, or with trophozoite cell lysis as described above for 3 h and 6 h at MOI1.2. The ELISAs were performed on spent media from infection samples and uninfected control samples at the respective time points. The medium was collected, centrifuged (1000 x g, 10 min, 4°C) to remove cellular debris and stored at -20°C until analysis. The assayed cytokines included CXCL1 and CXCL8. CXCL1 was measured using the Human CXCL1/GRO alpha Quantikine ELISA Kit (R&D Systems) and CXCL8 was measured using the IL-8 Human ELISA Kit (cat# KHC0081, Thermo Fisher [Invitrogen]) according to the manufacturer’s instructions. Absorbance reads from samples and standard curves were plotted in GraphPad Prism version 9.2.0, GraphPad Software, San Diego, California USA, www. graphpad.com. Cytokine concentrations were obtained by using a four-parameter logistic ELISA curve fitting. Statistical significance in protein concentrations was determined using a one-way analysis of variance (ANOVA) at α < 0.05, followed by Bonferroni comparisons (P < 0.05).

CXCL1 concentrations in conditioned media of hPSCs were measured using the Human CXCL1/GRO alpha Quantikine ELISA Kit (R&D Systems) according to the manufacturer’s instructions. The absorbance was measured at an optical density (OD) of 450 nm using Spectra Max M2e (Molecular Devices, Sunnyvale, CA, USA).