duo set elisa development kits  (R&D Systems)

Journal: The Journal of Obstetrics and Gynaecology Research

Article Title: Impact of tumor necrosis factor‐alpha and lysophosphatidic acid on the behavior of ovarian cancer cells in a three‐dimensional collagen hydrogel

doi: 10.1111/jog.70026

Figure Lengend Snippet: VEGF secretion in response to TNF‐α or LPA at two different concentrations. The concentration of VEGF was measured with ELISA in SKOV‐3 (a and b), OVCAR‐8 (c and d), OVCAR‐5 (e and f), and OVCAR‐4 (g and h) cell lines grown in collagen gels for 2–8 days with the appropriate concentration of vehicle (control gels), TNF‐α or LPA. Protein concentrations of VEGF accumulated over 48 h in the culture medium are shown as mean + SEM from ≥3 independent experiments. Statistical significance p * <0.05; ** <0.01; **** <0.0001. LPA, lysophosphatidic acid; TNF‐α, tumor necrosis factor‐alpha; VEGF, vascular endothelial growth factor.

Article Snippet: Protein levels were measured using the human VEGF, human IL‐8, or human IL‐6 using the appropriate Duo SET ELISA development kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer's protocols.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of Obstetrics and Gynaecology Research

Article Title: Impact of tumor necrosis factor‐alpha and lysophosphatidic acid on the behavior of ovarian cancer cells in a three‐dimensional collagen hydrogel

doi: 10.1111/jog.70026

Figure Lengend Snippet: IL‐8 secretion in response to TNF‐α or LPA at two different concentrations. The concentration of IL‐8 was measured with ELISA in SKOV‐3 (a and b), OVCAR‐8 (c and d), OVCAR‐5 (e and f), and OVCAR‐4 (g and h) cell lines grown in collagen gels for 2–8 days with the appropriate concentration of vehicle (control gels), TNF‐α or LPA. Protein concentrations of IL‐8 accumulated over 48 h in the culture medium are shown as mean + SEM from ≥3 independent experiments. Statistical significance p * <0.05; ** <0.01; *** <0.001; **** <0.0001. IL‐8, interleukin‐8; LPA, lysophosphatidic acid; TNF‐α, tumor necrosis factor‐alpha.

Article Snippet: Protein levels were measured using the human VEGF, human IL‐8, or human IL‐6 using the appropriate Duo SET ELISA development kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer's protocols.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of Obstetrics and Gynaecology Research

Article Title: Impact of tumor necrosis factor‐alpha and lysophosphatidic acid on the behavior of ovarian cancer cells in a three‐dimensional collagen hydrogel

doi: 10.1111/jog.70026

Figure Lengend Snippet: IL‐6 secretion from cells that were stimulated by two concentrations of TNF‐α or LPA. The concentration of IL‐6 was measured with ELISA in SKOV‐3 (a and b), OVCAR‐8 (c and d), OVCAR‐5 (e and f), and OVCAR‐4 (g and h) cell lines grown in collagen gels for 2–8 days with the appropriate concentration of vehicle (control gels), TNF‐α or LPA. Protein concentrations of IL‐6 accumulated over 48 h in the culture medium are shown as mean + SEM from ≥3 independent experiments. Note the different scales in the left versus right columns. Statistical significance * <0.05 ** <0.01 *** <0.001 **** <0.0001. Results for OVCAR‐5 with LPA (f) were below detection. IL‐6, interleukin‐6; LPA, lysophosphatidic acid; TNF‐α, tumor necrosis factor‐alpha.

Article Snippet: Protein levels were measured using the human VEGF, human IL‐8, or human IL‐6 using the appropriate Duo SET ELISA development kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer's protocols.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: Expression and release of CXCL12 by SMCs and EPCs. ( A ) Analysis of CXLC12 release using ELISA. Cell supernatants from monocultured SMCs and EPCs, treated as indicated, were collected 24 h after cultivation. * p < 0.05 vs. untreated cells; n = 6. ( B ) Real-time RT-PCR analysis of CXCL12 expression SMCs and EPCs treated as indicated. Results were normalized to CXCL12 expression in SMCs. * p < 0.05 vs. untreated cells; n = 5. ( C ) Detection of CXCL12 in MVs derived from monocultured SMCs and EPCs. Isolated MVs were lysed in RIPA buffer and CXCL12 levels were determined using ELISA. * p < 0.05 vs. non-injured SMCs, # p < 0.05 vs. SMC-MV; n = 5. ( D ) Enumeration of MVs in the supernatant of EPCs, non-injured SMCs and injured SMCs using flow cytometry with calibrated microbeads. * p < 0.05 vs. non-injured SMCs; n = 4. ( E ) Evaluation of the effect of EPC-SMC co-cultivation and engagement of CXCR4 on the release of CXCL12. Supernatants from monocultured SMCs, monocultured EPCs and EPCs co-cultured with SMCs, each in the presence or absence of a blocking CXCR4 Ab, were analyzed for CXCL12 concentration using ELISA. * p < 0.05 vs. SMCs, # p < 0.05 vs. EPC-SMC co-culture in the absence of anti-CXCR4; n = 5. ( F ) Real-time RT-PCR analysis of CXCL12 expression to test the impact of EPC-SMC co-cultivation and involvement of CXCR4. CXCL12 transcripts were determined in SMCs, EPCs and EPC-SMC co-cultures in the presence or absence of a blocking CXCR4 Ab. * p < 0.05 vs. SMCs, # p < 0.05 vs. EPC-SMC co-culture in the absence of anti-CXCR4; n = 5. ( G ) Real-time RT-PCR analysis of CXCL12 expression in SMCs treated with various doses of rCXCL12, CM-EPC or EPC-MV in the presence or absence of an anti-CXCR4 Ab. * p < 0.05 vs. untreated SMCs (control), # p < 0.05 vs. respective treatment in the absence of anti-CXCR4; n = 5. ( H ) Adhesion of EPCs to SMCs under flow conditions in vitro. EPCs pretreated with/without an anti-CXCR4 Ab were perfused in a parallel flow chamber and the number of cells EPCs adherent to the SMC monolayer was determined and expressed as adherent cells per 1 mm². For some experiments, the SMC monolayer was wounded by a linear scratch before perfusion of EPCs. * p < 0.05 vs. untreated and non-scratched SMCs (control), # p < 0.05 vs. respective treatment in the absence of anti-CXCR4; n = 4 to 6.

Article Snippet: Concentrations of CXCL12 and PDGF-BB were determined with the Duo-Set ELISA Development Kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Derivative Assay, Isolation, Flow Cytometry, Cell Culture, Blocking Assay, Concentration Assay, Co-Culture Assay, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: CXCL12–CXCR4-dependent control of SMC phenotype. ( A – E ) Analysis of EPC-mediated modulation of SMC phenotype and the involvement of the CXCL12–CXCR4 axis. SMCs were treated as indicated for 48 h and presence of SMA and Calponin was measured using flow cytometry. Data are expressed as mean fluorescence intensity (MFI) in % normalized to untreated SMCs (control). ( A – C ) * p < 0.05 vs. control and # p < 0.05 vs. SMCs treated with rCXCL2 50 ng/mL, ( D ) * p < 0.05 vs. SMCs co-cultured with EPCs, ( E ) * p < 0.05 vs. control and # p < 0.05 vs. the respective treatment of SMCs with CM-EPC or EPC-MV in the absence of anti-PDGFRß; n = 4 to 6. ( F ) Comparison of CXCL12 and PDGF-BB released by monocultured EPCs, monocultured SMCs and EPC-SMC co-cultures. Secreted paracrine factors present in the supernatant of the respective cultures were assessed using ELISA; n = 5.

Article Snippet: Concentrations of CXCL12 and PDGF-BB were determined with the Duo-Set ELISA Development Kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Flow Cytometry, Fluorescence, Cell Culture, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Anti-inflammatory and Anti-oxidative Effects of Phytohustil ® and Root Extract of Althaea officinalis L. on Macrophages in vitro

doi: 10.3389/fphar.2020.00290

Figure Lengend Snippet: Anti-inflammatory effects of Phytohustil ® (Phyto.), its excipients (Exp), or REAo (Extr.) against (A) LPS–induced Tumor Necrosis Factor (TNF-α) and (B) Interleukin-6 (IL6) release (measured by ELISA) in human MΦ compared to diclofenac sodium salt (Diclo) treatment. Medium control (med. control). Data are given as mean + SEM; ** p < 0.01, *** p < 0.001 (by T -TEST) significance vs. 3 h LPS treatment taken as 100%; + p < 0.05, +++ p < 0.001 vs. control and # p < 0.05, ## p < 0.01 vs. 200 μg/ml diclofenac. N = 5–7 independent experiments.

Article Snippet: Human TNF-α or IL6 were determined in the culture medium using the assay Duo Set ELISA Development kit (R&D Systems Europe, Ltd., Abingdon, United Kingdom) following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay