solid-phase sandwich enzyme-linked immunosorbent assay kit quantikins  (DPC Biermann GmbH)

Journal: Biology of reproduction

Article Title: Activin A promotes hyaluronan production and upregulates versican expression in human granulosa cells†.

doi: 10.1093/biolre/ioac070

Figure Lengend Snippet: Figure 7. The effects of the type I receptor inhibitor SB431542 on the activin A-induced increase in hyaluronan accumulation in SVOG and primary hGL cells. (A and C) SVOG (A) and primary hGL (C) cells were treated with 25 ng/mL and 50 ng/mL activin A for 12 h, respectively. The accumulated levels of hyaluronan in conditioned medium were measured using ELISA. (B and D) SVOG (B) and primary hGL (D) cells were pretreated with DMSO or SB431542 (10 μM) for 30 min, and the cells were treated with Ctrl or 25 ng/mL (SOVG hGL) and 50 ng/mL (primary hGL) activin A for an additional 12 h. The accumulated levels of hyaluronan in conditioned media were examined using ELISA. The results are expressed as the means ± SEM of at least three independent experiments. Student’s t-test was used to analyze two groups (∗P < 0.05). One-way ANOVA was used to analyze multiple groups (values without a common letter were significantly different, P < 0.05).

Article Snippet: After treatment, the accumulated levels of hyaluronan in culture medium were measured using a hyaluronan Quantikine solid phase sandwich enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Minneapolis, MN, DHYAL0).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression.

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Fig. 2 Upregulation of IL6 stimulated by E2 in NSCLC cell lines, A549 and H975. (a) ERβ/IL6 expression was up regulated in NSCLC cell lines compared with normal pneumocytes (HBE). (b) Synchronized cells were treated with E2 at different concentrations (0 nM, 1 nM, 10 nM and 100 nM) and Ful (0.1 μM and 1 μM) for 2 days. The protein expression of ERβ and IL6 was analyzed using western blot. (c) Synchronized cells were treated with E2 at different time points (0 h, 6 h, 12 h, 24 h, 48 h and 72 h) at concentrations of 10 nM. The protein expression of ERβ and IL6 was analyzed using western blot. GAPDH was used as a control. (d) Autocrine IL6 was analyzed by ELISA assay after concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM). (e) The upregulation of IL6 in A549 after stimulation with E2 (10 nM) or Ful (0.1uM) was determined by immunofluorescence

Article Snippet: IL-6 protein levels were detected in culture medium using solid phase sandwich enzyme-linked immunosorbent assay (ELISA) assays according to the manufacturer’s protocol (D6050, R&D Systems).

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Concentration Assay, Immunofluorescence

Journal: Journal of experimental & clinical cancer research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression.

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Fig. 3 Upregulation of IL6 stimulated by E2 treatment regulates aggressiveness of NSCLC cells. (a) The cell proliferation of A549 was determined by CCK-8 assay after 72 h concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM) or rhIL6 (0.5 ng/ml) or Stat3 inhibitor static (20uM). The results are representative of three independent experiments. (b) Colony formation assay measuring the proliferative activity in A549 cells after concentration-dependent treatment of E2 (10 nM) or Ful (0.1uM) or rhIL6 (0.5 ng/ml) or Stat3 inhibitor static (20uM). (c) Wound-healing assays were performed to assess NSCLC cell migration. Wound closure was determined 24 h after the scratch. (d) Transwell assay was used to quantify cell migration and invasion capacity. The average number of cells per field of view in three different experiments is plotted. (e-f) ELISA and western blot were used to detect the effect of E2 (10 nM) and its receptor antagonist Ful (0.1uM) on IL6 expression and the influence of MEK inhibitor U0126 (60 nM) or a selective PI3K inhibitor of LY294002 (0.6uM) on E2-mediated IL6 expression through MEK/ERK and PI3K/AKT activation in A549 cells

Article Snippet: IL-6 protein levels were detected in culture medium using solid phase sandwich enzyme-linked immunosorbent assay (ELISA) assays according to the manufacturer’s protocol (D6050, R&D Systems).

Techniques: CCK-8 Assay, Concentration Assay, Colony Assay, Activity Assay, Migration, Transwell Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Activation Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: 17β-estradiol upregulates IL6 expression through the ERβ pathway to promote lung adenocarcinoma progression.

doi: 10.1186/s13046-018-0804-5

Figure Lengend Snippet: Fig. 4 E2 regulates IL6 expression through ERβ and affects malignancy of lung cancer. (a) Autocrine IL6 was analyzed by ELISA assay after overexpression or knockdown of ERβ. (b) Upregulation of IL6 induced by E2 (10 nM) treatment 48 h was determined by immunofluorescence in A549. (c) Colony formation assay measuring the proliferative activity in A549 cells after overexpression or knockdown of ERβ. (d) Wound-healing assays were performed to assess NSCLC cell migration in response to changes in ERβ expression. (e-f) Transwell assay and DQ-collagen invasion assay were used to quantify the effect of ERβ regulation on cell migration and invasion capacity. (g) Expression of IL6 by western blotting after modulation of ERβ expression

Article Snippet: IL-6 protein levels were detected in culture medium using solid phase sandwich enzyme-linked immunosorbent assay (ELISA) assays according to the manufacturer’s protocol (D6050, R&D Systems).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Over Expression, Knockdown, Immunofluorescence, Colony Assay, Activity Assay, Migration, Transwell Assay, Invasion Assay, Western Blot